The forkhead box(FOX)family represents a class of transcription factors characterized by a distinctive winged helical structure.Forkhead box A1(FOXA1),a member of the forkhead box A(FOXA)subfamily within the FOX gene ...The forkhead box(FOX)family represents a class of transcription factors characterized by a distinctive winged helical structure.Forkhead box A1(FOXA1),a member of the forkhead box A(FOXA)subfamily within the FOX gene family,was the first forkhead protein identified in mammals.It serves as a pivotal transcription factor in tissue-specific differentiation and functions.Upon activation,owing to its unique structural domains,FOXA1 can interact with nucleosomes to open chromatin,thereby facilitating the recruitment of other transcription factors.These factorsmay act independently or synergistically with recruited transcription factors to regulate gene expression.Consequently,FOXA1 and other FOXA subfamily members with similar functions are referred to as“pioneer factors.”In recent years,studies on FOXA1 have advanced our understanding of its crucial role in gene regulation and involvement in disease processes.However,owing to their tissue-specific effects and varying biological behaviors in different environmental contexts,the underlying mechanisms remain elusive.Weused the PubMed database to better understand the complexmechanisms of FOXA1.By using keywords such as“FOXA1”and“transcription factor,”an extensive literature was retrieved,and many of the most relevant publications were screened.The selected studies were then thoroughly synthesized and summarized.This review synthesizes recent findings on FOXA1,encompassing its structural characteristics,domain functions,roles in embryonic development and the maintenance of adult organ morphology and function,interactions with histone posttranslational modifications in gene regulation,and the influence of its posttranslational modifications on gene expression.We also explore the involvement of FOXA1 in various diseases.By elucidating the biological mechanisms and disease-related roles of FOXA1,this review aims to provide insights for future research on its complex mechanisms and potential therapeutic targets.展开更多
BACKGROUND ANAPC1,a key regulator of the ubiquitination in tumour development,has not been thoroughly studied in hepatocellular carcinoma(HCC).AIM To elucidate the expression of ANAPC1 in HCC and its potential regulat...BACKGROUND ANAPC1,a key regulator of the ubiquitination in tumour development,has not been thoroughly studied in hepatocellular carcinoma(HCC).AIM To elucidate the expression of ANAPC1 in HCC and its potential regulatory mechanism related to ubiquitination.METHODS Bulk RNA(RNA sequencing and microarrays),immunohistochemistry(IHC)tissues,and single-cell RNA sequencing(scRNA-seq)data were integrated to comprehensively investigate ANAPC1 expression in HCC.Clustered regularly interspaced short palindromic repeats analysis was performed to assess growth in HCC cell lines following ANAPC1 knockout.Enrichment analyses were conducted to explore the functions of ANAPC1.ScRNA-seq data was used to examine the cell cycle and metabolic levels.CellChat analysis was applied to investigate the interactions between ANAPC1 and different cell types.The relationship between ANAPC1 expression and drug concentration was analyzed.RESULTS ANAPC1 messenger RNA was found to be upregulated in bulk RNA,IHC tissues samples and malignant hepatocytes.The proliferation of JHH2 cell lines was most significantly inhibited after ANAPC1 knockdown.In biological pathways,the development of HCC was found to be linked to the regulation of ubiquitin-mediated proteolysis.Additionally,scRNA-seq results indicated that highly expressed ANAPC1 was in the G2/M phase,with increased glycolysis/gluconeogenesis activity.A CellChat analysis showed that ANAPC1 was associated with the regulation of the migration inhibitory factor-(cluster of differentiation 74+C-X-C chemokine receptor type 4)pathway.Higher ANAPC1 expression correlated with stronger effects of sorafenib,dasatinib,ibrutinib,lapatinib,nilotinib and afatinib.CONCLUSION The high expression level of ANAPC1 may regulate the cell cycle and metabolic levels of HCC through the ubiquitination-related pathway,thereby promoting disease progression.展开更多
This study is based on wireless optogenetic technology,utilizing the CRY2/CIB1 photosensitive system to achieve spatiotemporal control of PD-L1 expression.In vitro experiments showed that the surface PD-L1 positivity ...This study is based on wireless optogenetic technology,utilizing the CRY2/CIB1 photosensitive system to achieve spatiotemporal control of PD-L1 expression.In vitro experiments showed that the surface PD-L1 positivity rate of cells increased from 28.6±3.1%to 67.3±5.4%(P<0.001).In animal experiments,the terminal tumor volume in the light exposure group was 450±90 mm3,with a tumor inhibition rate of approximately 49.4%(P<0.001),and the median survival was extended to 32 days(compared to 24 days in the control group,P=0.004).Immunological tests revealed a significant increase in CD8+T cell infiltration(112±18 vs 52±10 cells/HPF,P<0.01),a 30%decrease in the proportion of Tregs(P<0.05),and an increase in the M1/M2 macrophage ratio to 1.8.The results suggest that the wireless optogenetic system can not only precisely regulate PD-L1 but also remodel the tumor immune microenvironment,providing a new approach for precise immunotherapy of GBM.展开更多
To investigate the expression characteristics of sex related gene of FTZ-F 1 in the half-smooth tongue-sole （Cynoglossus semilaevis）, the homologue FTZ-F1 （hsFTZ-F1） full-length cDNA was isolated from the testis b...To investigate the expression characteristics of sex related gene of FTZ-F 1 in the half-smooth tongue-sole （Cynoglossus semilaevis）, the homologue FTZ-F1 （hsFTZ-F1） full-length cDNA was isolated from the testis by homologous cloning, and the cDNA included the open reading frame and a 66bp 5＇-UTR, along with a 1619bp 3＇-UTR, encoding a predicted 485 amino acid protein. Sequence, tissue distribution and phylogenic analyses of the FTZ-F1 showed that the hsFTZ-F1 belonged to SF-1/Ad4BP group. The hsFTZ-F1 transcripts were highly abundant in the gonads, kidneys, brain and head-kidneys, but weakly in other tissues. However, the expression level in the brain and head-kidney of female was highly abundant than in the male. The hsFTZ-F1 expression was highly abundant in the embryo than in the larvae, which suggested that the hsFTZ-F1 may be involved in the organogenesis in the tongue sole.展开更多
[ Objective] The aim of this study was to investigate effects of various inducers on the expression of cytochrome P450 CYP305 B1 V1 Gene in different tissues of wild mulberry silkworm. [ Method] Referring to the mRNA ...[ Objective] The aim of this study was to investigate effects of various inducers on the expression of cytochrome P450 CYP305 B1 V1 Gene in different tissues of wild mulberry silkworm. [ Method] Referring to the mRNA sequence of CYP305 B1 V1 Gene published in GenBank for wild mulberry silkworm, one pair of primers was designed, and the expression of cytochrome P450 CYP305 B1 V1 Gene in different tissues of wild mulberry silkworm treated by NaF, rutin, cypermethrin and ecdysone was also analyzed by the semi - quantitative RT - PCR. Furthermore, homology comparison and phylogenetic analysis for amino acid sequences of this gene were studied. [ Result] Rutin, cypermethrin and NaF had effects on the expression of P450 CYP305 B1 V1 Gene in different tissues of wild mulberry silkworm, while ecdysone had no significant effect. Homology comparison for amino acids indicated that the amino acid sequence of this gene was the most similar to that of CYP305 B1 gene in Bombyx mori with 100% amino acid identity, and highly similar to those of Tribolium casmneum CYP305A1, Apis mellifera CYP305A1, Drosophi- la melanogaster CYP305A1, Anopheles gambiae CYP305A2and Culex pipiens quinquefasciatus CYP2LI. [ Conclusion] CYP305 B1 V1 Gene of wild mulberry silkworm is likely to mainly take part in the metabolism of exogenous compounds, which is of great significance for revealing the function of cytochrome P450 and the metabolic mechanism of different drugs.展开更多
[Objective] The research aimed to induce the expression of FMDV structural protein VP1 in E.coli and purify the protein,then detect the activity.[Method] The fragment coding VP1 was amplified by PCR and doubly digeste...[Objective] The research aimed to induce the expression of FMDV structural protein VP1 in E.coli and purify the protein,then detect the activity.[Method] The fragment coding VP1 was amplified by PCR and doubly digested with BamH Ⅰ and XhoⅠ,then cloned into expression vector pGEX-4T-1 and pPROExHTb respectively to get recombinant plasmid pGEX-4T-1-VP1 and pPROExHTb-VP1.The recombinant plasmid pGEX-4T-1-VP1 and pPROExHTb-VP1 was transformed into E.coli BL21(DE3)and induced by IPTG,fusion protein was identifie...展开更多
According to the complete genome of foot-and-mouth disease virus(FMDV)type O,a pair of special primers was designed to amplify VP1 gene.The VP1 gene was amplified by RT-PCR and subsequently inserted into the expressio...According to the complete genome of foot-and-mouth disease virus(FMDV)type O,a pair of special primers was designed to amplify VP1 gene.The VP1 gene was amplified by RT-PCR and subsequently inserted into the expression vector pGEX-6p-1 and induced by IPTG.Then SDS-PAGE showed the expressed protein was 51 kD in molecular weight.Then the product was purified by GSTrap FF columns.The product was detected through Western-blot that showed the protein has antigenicity.It provided fundamental data and materials for further investigation on diagnosis method of FMDV.展开更多
[Objective]The aim was to explore the molecular mechanism of plant resistance to various stress response.[Method]The expression of LeWRKY1 in tomato seedlings under treatment with B.cinerea,exogenous JA and SA were ex...[Objective]The aim was to explore the molecular mechanism of plant resistance to various stress response.[Method]The expression of LeWRKY1 in tomato seedlings under treatment with B.cinerea,exogenous JA and SA were explored by real time quantitative RT-PCR technology.[Result]JA induced the expression of LeWRKY1,but SA did not.LeWRKY1 expression was up-regulated under B.cinerea infection.[Conclusion]LeWRKY1 might be involved in the tomato defense response to B.cinerea through JA dependent but SA independent signal pathway.展开更多
Differences of gene expression between salinity_stressed and control rice ( Oryza sativa L. ssp. indica ) cultivar “Zhaiyeqing 8' were compared using differential display PCR (DD_PCR) technique. Sequence an...Differences of gene expression between salinity_stressed and control rice ( Oryza sativa L. ssp. indica ) cultivar “Zhaiyeqing 8' were compared using differential display PCR (DD_PCR) technique. Sequence analysis of one salt_inducible cDNA clone revealed that this clone represented a new member of rice translation elongation factor 1A (eEF1A) gene family and was tentatively named REF1A. Northern blot hybridization using REF1A fragment as a probe was performed to investigate the expression of rice translation elongation factor 1A gene in response to various environmental factors. It was observed that expression of the eEF1A gene in rice shoots was dramatically induced by salinity stress or exogenous application of abscisic acid (ABA). The induction of this gene by ABA stress occurred more quickly than that by salinity stress. In addition, expression of rice translation elongation factor 1A gene was also induced by drought (15% PEG6000), cold (4 ℃) or heat_shock (37 ℃) stresses. The results suggested that the induction of translation elongation factor 1A gene expression by environmental stresses might reflect the general adaptive response of rice plants to the adverse circumstances.展开更多
We investigated the inheritance and expression of cry1A gene in transgenic maize ( Zea mays L.) by Southern blotting analysis and enzyme_linked immunosorbent assays (ELISA). The results showed that cry1A ha...We investigated the inheritance and expression of cry1A gene in transgenic maize ( Zea mays L.) by Southern blotting analysis and enzyme_linked immunosorbent assays (ELISA). The results showed that cry1A had been transmitted to progeny of transgenic maize as a single gene. Contents of cry1A insecticidal protein were significantly different among transgenic maize lines and various tissues of the same transgenic lines. High expression of cry1A protein occurred in green tissues, such as leaf and husk leaf, and low expression occurred in pith, tassel, ear pith, pollen and silk. The results also showed that the contents of cry1A insecticidal protein in leaves of transgenic maize increased with the advance of development and there was no significant difference in cry1A expression level among various generations of transgenic maize.展开更多
[Objective] The research aimed to provide the theoretical basis for establishing a rapid diagnosis method for porcine parvovirus（PPV）. [ Method] One pair of primers were designed according to PPV genome sequences on...[Objective] The research aimed to provide the theoretical basis for establishing a rapid diagnosis method for porcine parvovirus（PPV）. [ Method] One pair of primers were designed according to PPV genome sequences on GenBank website and the sequences of prokaryotic expression vector pET30a （ ＋ ） with multiple cloning sites. The whole sequence of NS1 gene in PPV SD1 strain was amplified by using PCR technology and the positive recombinant plasmid was analyzed by sequencing and homology comparison. The prokaryotic expression recombinant plasmid PET30a/NS1 was constructed to make its induction expression in Escherichia coll. [ Result] The target fragment with the length of 2 208 bp was obtained from PCR amplification. The nucleotide homologies between the cloned NS1 gene and the reported relevant PPV genes were from 97.3 % to 99.4 %, which indicated that NS1 gene had high conservation. But it had a 12-basepair successive deletion near the hydroxyl end. The cloned PPV NS1 gene was successfully expressed in prokaryotic cell, and its expression products existed mostly in inclusion bodies. [ Conclusion] The results of SDS-PAGE detection showed that the molecular weight of PPV NS1 protein was 86 KD.展开更多
[Objective] To construct prokaryotic expression vectors encoding gene Erb3binding protein （EBP1）, which plays important roles in regulating plant organ size from Nervilia fordii （Hance） Schltr. [Methods] PCR produ...[Objective] To construct prokaryotic expression vectors encoding gene Erb3binding protein （EBP1）, which plays important roles in regulating plant organ size from Nervilia fordii （Hance） Schltr. [Methods] PCR products of NfEBP1 with particular restriction sites and expression vectors, pET-28 and pET-16b were digested. Ligation, transformation and selection were performed to construct the recombinant plasmids pET-28-NfEBP1 and pET-16-NfEBP1. The recombinant plasmids were transformed into E. coli BL21 using heat -shock transformation. [Results] Recombinant plasmids pET-28-NfEBP1-1188 and pET-16-NfEBP1-1188 were constructed and transformed into expressional host cells, E. coli BL21, and validated by colony PCR, sequencing and double digestion. [Conclusion] Prokaryotic expression vectors of EBP1 gene from N. fordii were successfully constructed, which laid the foundation for characterization of the gene function.展开更多
[ Objective ] This study was to express and purify Arabidopsis thaliana heat shock factor HSF1. [ Method ] Using Escherichia coli M15 harboring HSF1 （pQE32/His6-HSF1, pREP4） as experimental materials, HSF1 was induc...[ Objective ] This study was to express and purify Arabidopsis thaliana heat shock factor HSF1. [ Method ] Using Escherichia coli M15 harboring HSF1 （pQE32/His6-HSF1, pREP4） as experimental materials, HSF1 was induced to express with isopropyl-β-D-galactoside （IPTG） ; then the expression product was purified using Ni-NTA-agarose affinity chromatography and analyzed by SDS-PAGE. [Result] HSF1 of Arabidopsis thaliana was successfully expressed and purified. [ Conclusion] This study provides materials for understanding the blinding site of HSF1 on Arabidopsis thaliana chromosome, further laying a good foundation for revealing the regulatory mechanism and physiological function of HSF1.展开更多
Zymography and in situ hybridization were used to investigate matrixmetalloproteinase -2, -9 (MMP -2, MMP-9) activities and expressions of MMP -2, -9 and TIMP1, -2, -3 (tissue inhibitors of matrix metallo-proteinases)...Zymography and in situ hybridization were used to investigate matrixmetalloproteinase -2, -9 (MMP -2, MMP-9) activities and expressions of MMP -2, -9 and TIMP1, -2, -3 (tissue inhibitors of matrix metallo-proteinases) mRNA in the rat uterus during estrouscycle. The relative activity was semiquanted by using densitometric analysis. The MMP-2(67 kDa) activity in every stage during estrpus cycle was detected by zymography. MMP-2activity was highest at proestrus; higher at estrus and metaestrus; lowest at diestrus. Throughin situ hybridization, MMP -2, -9, TIMP -1~ -3 mRNA mainly in hasal stroma cells of uterineendometrium were detected. The positive signals of MMP -2 and -9 mRNAs in hasal stromacells were shown stronger at proestrus, estrus and metaestrus while they showed the weakest atdiestrus. The expression of MMP -2 mRNA coincided with MMP -2 activity change. MMP-2and -9 mRNAs were also highly expressed in uterine circular muscle at estrus. Weak signals ofMMP -9 mRNA were detected in uterine luminal and glandular epithelial cells at estrus.TIMP -1 mRNA in hasal stroma cells was shown as the strongest expression at estrus andmetaestrus; stronger at proestrus and the weakest at diestrus. TIMP-2 mRNA in basal stromacells was stronger at estrus and diestrus; weaker at proestrus and metaestrus. TIMP -1 and -2mRNAs were also highly expressed in uterine luminal and glandular epithelial cells at estrus.TIMP -3 mRNA in hasal stroma cells revealed the strongest expression at estrus; stronger atdiestrus and metaestrus and showed the weakest at proestrus. The mRNA was also highlyexpressed in uterine circular muscle at estrus. In short, our present results provide evidencethat MMP -2, -9 and TIMP -1~ -3 were involved in rat uterine endometrium reconstructionduring estrous cycle.展开更多
[Objective] The aim of experiment was to lay molecular foundation for studying maturity mechanism of banana after harvest. [Method] The combined method of suppressing subtractive hybridization and cDNA micro-array wer...[Objective] The aim of experiment was to lay molecular foundation for studying maturity mechanism of banana after harvest. [Method] The combined method of suppressing subtractive hybridization and cDNA micro-array were used to obtain cDNA segment of one PRMT gene in banana and the whole cDNA sequence of the gene was cloned.The bioinformatics analysis was operated on it,in addition, the expression profile analysis was conducted in different organs and different mature periods of banana.[Result] The whole length of cDNA in MaPRMT1 was 1 158 bp and possessed a complete open reading frame,which could encode 385 amino acids.It had high homology with PRMT in plant,containing one Methyltransf_1 domain.The MaPRMT1 gene was expressed in root,stem,leaf and fruit of banana and the expression levels in stem and leaf were relatively high.As the increase of days after harvest,the expression level declined gradually,however it reached maximum when ethylene release was biggest,then it declined.[Conclusion] MaPRMT1 belonged to the first kind of arginine methyltransferase and it was expressed differently in different organs and fruits at different mature periods.展开更多
A genomic DNA fragment containing the 5'-upstream sequence and part of the open reading frame corresponding to the cytosolic fructose-1,6-bisphosphatase (cyFBPase) cDNA was isolated by Genome Walking. The 1 195 li...A genomic DNA fragment containing the 5'-upstream sequence and part of the open reading frame corresponding to the cytosolic fructose-1,6-bisphosphatase (cyFBPase) cDNA was isolated by Genome Walking. The 1 195 lip 5'-flanking region which started from the translation initiation ATG codon was fused to reporter gene encoding beta-glucuronidase (GUS) and stably transferred to rice via particle bombardment. Strong GUS activity was detected in leaves and leaf sheaths of transgenic rice, but not in culms and roots. Histochemical localization revealed that GUS expression was exclusively restricted to mesophyll cells in transgenic rice. Our results indicate that the 1 195 bp fragment contains all the cis-elements required for directing mesophyll-specific expression pattern in rice.展开更多
We describe the temporal and spatial expression pattern of Sox 1 gene during Xenopus laevis early development and compare the expression patterns of Sox 1-3 in the developing eye and brain. Alignment of Sox 1-3 amino ...We describe the temporal and spatial expression pattern of Sox 1 gene during Xenopus laevis early development and compare the expression patterns of Sox 1-3 in the developing eye and brain. Alignment of Sox 1-3 amino acid sequences shows a high conservation within the HMG-box DNA binding domains. RT-PCR analysis indicates that Sox 1 is expressed throughout development from the unfertilized egg to at least the tadpole stage, although at different expression levels. The transcripts of XSox 1 are detected in the animal pole at cleavage and blastrula stages and mainly in the central nervous system （CNS） and the developing eye at neurula stages. The study of the developmental expression of XSox 1 will aid in the elucidation of the function of SoxB 1 subgroup genes in vertebrate neurogenesis.展开更多
[Objective] This study was conducted to investigate cloning and expression of Pun1 gene controlling pungency of pepper. [Method] With Capsicum annuum L as a material, the cDNA sequence of Capunl gene was obtained, wit...[Objective] This study was conducted to investigate cloning and expression of Pun1 gene controlling pungency of pepper. [Method] With Capsicum annuum L as a material, the cDNA sequence of Capunl gene was obtained, with a total length of 1 457 bp, coding 440 amino acids. [Result] Phylogenetic analysis showed that Capunl was closest to Pun1 of C. chinense, with a genetic distance of 0.019 3. Plant expression vector pCAM-Punl-GFP was constructed and transformed into to- bacco, and it was found that the protein coded by fusion gene Punl：：GFP was lo- cated on cell membrane. Prokaryotic expression vectors were constructed, and by SDS-PAGE and Western Blot detection, an induced protein with a molecular weight of 63 ku was obtained. It was found by real-time fluorescence quantitative expres- sion that Pun1 gene was expressed at the highest level 30 d after flowering, de- creased then, and could not be detected substantially 40 and 45 d after flowering. [Conclusion] This study provides information and reference for molecular regulation mechanism of Pun1 gene.展开更多
The Raphanus sativus L. antifungal protein 1 (Rs_AFP1) gene was isolated by polymerase chain reaction (PCR). The complete open reading frame and the fragment encoding the putative mature protein were inserted into the...The Raphanus sativus L. antifungal protein 1 (Rs_AFP1) gene was isolated by polymerase chain reaction (PCR). The complete open reading frame and the fragment encoding the putative mature protein were inserted into the prokaryotic expression vector pET_32b(+), respectively. Subsequent expression showed that the Rs_AFP1 was produced in E. coli as a 27 kD fusion protein only when the N_terminal signal peptide was removed. After treatment with thrombin to remove part of the N_terminal His.tag sequence, the bacterially expressed Rs_AFP1 was used for fungal growth inhibition assay which was conducted on Verticillium dahliae Kleb., a soil_born fungus causing the cotton wilt disease. Results showed that, in the liquid medium, the Rs_AFP1 fusion protein at a concentration of 0.3 g/L clearly inhibited the growth of V. dahliae and the germination of spores. Thus the bacterially expressed fusion protein had the antifungal activity against V. dahliae.展开更多
AIM To detect the congenital expression patterns of mdr 1 gene in commonly encountered malignant tumors in clinic, and the relationship between the expression of mdr 1 gene and the prognostic morphology in ...AIM To detect the congenital expression patterns of mdr 1 gene in commonly encountered malignant tumors in clinic, and the relationship between the expression of mdr 1 gene and the prognostic morphology in esophageal carcinomas. METHODS A total of 151 resected samples of malignant tumors without preoperative treatment were taken from Anyang City Tumor Hospital. The congenital expression of their mdr 1 gene was detected with reverse transcription polymerase chain reaction (RT PCR) and was compared with each other. The positive incidence of mdr 1 gene in 46 samples of esophageal carcinoma was compared with their differentiated grades, TNM stages and macroscopic types, and the precautions and advantages of RT PCR were evaluated. RESULTS All the 151 samples were confirmed to be malignant histopathologically, including cancers of stomach and gastric cardia (n =51), esophagus ( n =46), colorectum ( n =16), breast ( n =15), thyroid ( n =10), lung ( n =9), uterine cervix ( n =24). The positive expression rate of their mdr 1 gene was 33 3%, 37%, 31 3%, 13 2%, 40%, 55%, and 0% respectively. All the 46 samples of esophageal carcinoma were pathologically confirmed to be squamous cell carcinoma. The total expression rate of their mdr 1 gene was 37% (17/46), 35% (6/17), 40% (8/20), and 33% (3/9) for differentiation grade Ⅰ, Ⅱ and Ⅲ respectively. The expression rate of TNM classification was 33% (6/18), 40% (5/12) and 37% (6/16) in stage Ⅱa, Ⅱb and Ⅲ. The expression rate was 33% (3/9) in ulcerous type, 37% (3/8) in constrictive one, 33% (5/15) in fungoid one, and 40% (6/14) in medullary one. No statistically significant difference was found. CONCLUSION Compared with other methods, RT PCR is more simple, reliable and accurate in detecting mdr 1 gene expression in tissues of tumor. The overexpression of mdr 1 gene in these neoplasms suggested that cases should be handled differently for chemotherapy with rational use of drugs. Excision is the chief treatment for carcinoma of esophagus. The expression of mdr 1 gene in tissues of esophageal cancer is correlated with the parameters of tumor molecular biology which are independent of histopathological morphology.展开更多
基金supported by grants from the National Natural Science Foundation of China(No.82470042)Liaoning Provincial Joint Science and Technology Plan(No.2023JH2/101800021)+4 种基金Basic Scientific Research Project of Liaoning Provincial Department of Education(No.LJKMZ20221186)ShenyangMunicipal PublicHealth Research and Development Special Project(No.LJKMZ20221186).The funders had no role in the design of the studyin the collection,analyses,or interpretation of datain the writing of the manuscriptor in the decision to publish the results.
文摘The forkhead box(FOX)family represents a class of transcription factors characterized by a distinctive winged helical structure.Forkhead box A1(FOXA1),a member of the forkhead box A(FOXA)subfamily within the FOX gene family,was the first forkhead protein identified in mammals.It serves as a pivotal transcription factor in tissue-specific differentiation and functions.Upon activation,owing to its unique structural domains,FOXA1 can interact with nucleosomes to open chromatin,thereby facilitating the recruitment of other transcription factors.These factorsmay act independently or synergistically with recruited transcription factors to regulate gene expression.Consequently,FOXA1 and other FOXA subfamily members with similar functions are referred to as“pioneer factors.”In recent years,studies on FOXA1 have advanced our understanding of its crucial role in gene regulation and involvement in disease processes.However,owing to their tissue-specific effects and varying biological behaviors in different environmental contexts,the underlying mechanisms remain elusive.Weused the PubMed database to better understand the complexmechanisms of FOXA1.By using keywords such as“FOXA1”and“transcription factor,”an extensive literature was retrieved,and many of the most relevant publications were screened.The selected studies were then thoroughly synthesized and summarized.This review synthesizes recent findings on FOXA1,encompassing its structural characteristics,domain functions,roles in embryonic development and the maintenance of adult organ morphology and function,interactions with histone posttranslational modifications in gene regulation,and the influence of its posttranslational modifications on gene expression.We also explore the involvement of FOXA1 in various diseases.By elucidating the biological mechanisms and disease-related roles of FOXA1,this review aims to provide insights for future research on its complex mechanisms and potential therapeutic targets.
基金Co-first authors:Yu-Xing Tang 0000-0003-4382-4942Co-first authors:Wei-Zi Wu+8 种基金Corresponding author:Gang Chen,MD,Professor,Department of Pathology,The First Affiliated Hospital of Guangxi Medical University,No.6 Shuangyong Road,Nanning 530021,Guangxi Zhuang Autonomous Region,China.chengang@gxmu.edu.cn,0000-0003-2402-2987Co-corresponding authors:Yan-Ting ZhanSheng-Sheng Zhou,0000-0003-2414-460XDa-Tong Zeng,0000-0002-3338-4122Guang-Cai Zheng,0009-0001-5921-6688Rong-Quan He,0000-0002-7752-2080Di-Yuan Qin,0009-0003-3214-4762Wan-Ying Huang,0000-0002-8314-5963Yu-Lu Tang,0009-0004-0462-618X。
文摘BACKGROUND ANAPC1,a key regulator of the ubiquitination in tumour development,has not been thoroughly studied in hepatocellular carcinoma(HCC).AIM To elucidate the expression of ANAPC1 in HCC and its potential regulatory mechanism related to ubiquitination.METHODS Bulk RNA(RNA sequencing and microarrays),immunohistochemistry(IHC)tissues,and single-cell RNA sequencing(scRNA-seq)data were integrated to comprehensively investigate ANAPC1 expression in HCC.Clustered regularly interspaced short palindromic repeats analysis was performed to assess growth in HCC cell lines following ANAPC1 knockout.Enrichment analyses were conducted to explore the functions of ANAPC1.ScRNA-seq data was used to examine the cell cycle and metabolic levels.CellChat analysis was applied to investigate the interactions between ANAPC1 and different cell types.The relationship between ANAPC1 expression and drug concentration was analyzed.RESULTS ANAPC1 messenger RNA was found to be upregulated in bulk RNA,IHC tissues samples and malignant hepatocytes.The proliferation of JHH2 cell lines was most significantly inhibited after ANAPC1 knockdown.In biological pathways,the development of HCC was found to be linked to the regulation of ubiquitin-mediated proteolysis.Additionally,scRNA-seq results indicated that highly expressed ANAPC1 was in the G2/M phase,with increased glycolysis/gluconeogenesis activity.A CellChat analysis showed that ANAPC1 was associated with the regulation of the migration inhibitory factor-(cluster of differentiation 74+C-X-C chemokine receptor type 4)pathway.Higher ANAPC1 expression correlated with stronger effects of sorafenib,dasatinib,ibrutinib,lapatinib,nilotinib and afatinib.CONCLUSION The high expression level of ANAPC1 may regulate the cell cycle and metabolic levels of HCC through the ubiquitination-related pathway,thereby promoting disease progression.
文摘This study is based on wireless optogenetic technology,utilizing the CRY2/CIB1 photosensitive system to achieve spatiotemporal control of PD-L1 expression.In vitro experiments showed that the surface PD-L1 positivity rate of cells increased from 28.6±3.1%to 67.3±5.4%(P<0.001).In animal experiments,the terminal tumor volume in the light exposure group was 450±90 mm3,with a tumor inhibition rate of approximately 49.4%(P<0.001),and the median survival was extended to 32 days(compared to 24 days in the control group,P=0.004).Immunological tests revealed a significant increase in CD8+T cell infiltration(112±18 vs 52±10 cells/HPF,P<0.01),a 30%decrease in the proportion of Tregs(P<0.05),and an increase in the M1/M2 macrophage ratio to 1.8.The results suggest that the wireless optogenetic system can not only precisely regulate PD-L1 but also remodel the tumor immune microenvironment,providing a new approach for precise immunotherapy of GBM.
基金Supported by grants from State 863 High-Technology R&D Project of China(2006AA10A403)Shandong Genetic Improvement Key Project for Agricultural OrganismDoctor Initial Funding of Guangdong Ocean University(0712103)
文摘To investigate the expression characteristics of sex related gene of FTZ-F 1 in the half-smooth tongue-sole （Cynoglossus semilaevis）, the homologue FTZ-F1 （hsFTZ-F1） full-length cDNA was isolated from the testis by homologous cloning, and the cDNA included the open reading frame and a 66bp 5＇-UTR, along with a 1619bp 3＇-UTR, encoding a predicted 485 amino acid protein. Sequence, tissue distribution and phylogenic analyses of the FTZ-F1 showed that the hsFTZ-F1 belonged to SF-1/Ad4BP group. The hsFTZ-F1 transcripts were highly abundant in the gonads, kidneys, brain and head-kidneys, but weakly in other tissues. However, the expression level in the brain and head-kidney of female was highly abundant than in the male. The hsFTZ-F1 expression was highly abundant in the embryo than in the larvae, which suggested that the hsFTZ-F1 may be involved in the organogenesis in the tongue sole.
文摘[ Objective] The aim of this study was to investigate effects of various inducers on the expression of cytochrome P450 CYP305 B1 V1 Gene in different tissues of wild mulberry silkworm. [ Method] Referring to the mRNA sequence of CYP305 B1 V1 Gene published in GenBank for wild mulberry silkworm, one pair of primers was designed, and the expression of cytochrome P450 CYP305 B1 V1 Gene in different tissues of wild mulberry silkworm treated by NaF, rutin, cypermethrin and ecdysone was also analyzed by the semi - quantitative RT - PCR. Furthermore, homology comparison and phylogenetic analysis for amino acid sequences of this gene were studied. [ Result] Rutin, cypermethrin and NaF had effects on the expression of P450 CYP305 B1 V1 Gene in different tissues of wild mulberry silkworm, while ecdysone had no significant effect. Homology comparison for amino acids indicated that the amino acid sequence of this gene was the most similar to that of CYP305 B1 gene in Bombyx mori with 100% amino acid identity, and highly similar to those of Tribolium casmneum CYP305A1, Apis mellifera CYP305A1, Drosophi- la melanogaster CYP305A1, Anopheles gambiae CYP305A2and Culex pipiens quinquefasciatus CYP2LI. [ Conclusion] CYP305 B1 V1 Gene of wild mulberry silkworm is likely to mainly take part in the metabolism of exogenous compounds, which is of great significance for revealing the function of cytochrome P450 and the metabolic mechanism of different drugs.
基金Supported by National Supporting Plan(2006BAD06A14)Transgenic Major Projects(2008ZX08011-004)~~
文摘[Objective] The research aimed to induce the expression of FMDV structural protein VP1 in E.coli and purify the protein,then detect the activity.[Method] The fragment coding VP1 was amplified by PCR and doubly digested with BamH Ⅰ and XhoⅠ,then cloned into expression vector pGEX-4T-1 and pPROExHTb respectively to get recombinant plasmid pGEX-4T-1-VP1 and pPROExHTb-VP1.The recombinant plasmid pGEX-4T-1-VP1 and pPROExHTb-VP1 was transformed into E.coli BL21(DE3)and induced by IPTG,fusion protein was identifie...
基金Supported by the Science and Technology Foundation from Science&Technology Department of Guangxi Autonomous Region(0779001)~~
文摘According to the complete genome of foot-and-mouth disease virus(FMDV)type O,a pair of special primers was designed to amplify VP1 gene.The VP1 gene was amplified by RT-PCR and subsequently inserted into the expression vector pGEX-6p-1 and induced by IPTG.Then SDS-PAGE showed the expressed protein was 51 kD in molecular weight.Then the product was purified by GSTrap FF columns.The product was detected through Western-blot that showed the protein has antigenicity.It provided fundamental data and materials for further investigation on diagnosis method of FMDV.
基金Supported by Beijing Nature Science Foundation(5102015)~~
文摘[Objective]The aim was to explore the molecular mechanism of plant resistance to various stress response.[Method]The expression of LeWRKY1 in tomato seedlings under treatment with B.cinerea,exogenous JA and SA were explored by real time quantitative RT-PCR technology.[Result]JA induced the expression of LeWRKY1,but SA did not.LeWRKY1 expression was up-regulated under B.cinerea infection.[Conclusion]LeWRKY1 might be involved in the tomato defense response to B.cinerea through JA dependent but SA independent signal pathway.
文摘Differences of gene expression between salinity_stressed and control rice ( Oryza sativa L. ssp. indica ) cultivar “Zhaiyeqing 8' were compared using differential display PCR (DD_PCR) technique. Sequence analysis of one salt_inducible cDNA clone revealed that this clone represented a new member of rice translation elongation factor 1A (eEF1A) gene family and was tentatively named REF1A. Northern blot hybridization using REF1A fragment as a probe was performed to investigate the expression of rice translation elongation factor 1A gene in response to various environmental factors. It was observed that expression of the eEF1A gene in rice shoots was dramatically induced by salinity stress or exogenous application of abscisic acid (ABA). The induction of this gene by ABA stress occurred more quickly than that by salinity stress. In addition, expression of rice translation elongation factor 1A gene was also induced by drought (15% PEG6000), cold (4 ℃) or heat_shock (37 ℃) stresses. The results suggested that the induction of translation elongation factor 1A gene expression by environmental stresses might reflect the general adaptive response of rice plants to the adverse circumstances.
文摘We investigated the inheritance and expression of cry1A gene in transgenic maize ( Zea mays L.) by Southern blotting analysis and enzyme_linked immunosorbent assays (ELISA). The results showed that cry1A had been transmitted to progeny of transgenic maize as a single gene. Contents of cry1A insecticidal protein were significantly different among transgenic maize lines and various tissues of the same transgenic lines. High expression of cry1A protein occurred in green tissues, such as leaf and husk leaf, and low expression occurred in pith, tassel, ear pith, pollen and silk. The results also showed that the contents of cry1A insecticidal protein in leaves of transgenic maize increased with the advance of development and there was no significant difference in cry1A expression level among various generations of transgenic maize.
基金Supported by the Natural Science Foundation Program of Shandong Province(2007ZRA16001)~~
文摘[Objective] The research aimed to provide the theoretical basis for establishing a rapid diagnosis method for porcine parvovirus（PPV）. [ Method] One pair of primers were designed according to PPV genome sequences on GenBank website and the sequences of prokaryotic expression vector pET30a （ ＋ ） with multiple cloning sites. The whole sequence of NS1 gene in PPV SD1 strain was amplified by using PCR technology and the positive recombinant plasmid was analyzed by sequencing and homology comparison. The prokaryotic expression recombinant plasmid PET30a/NS1 was constructed to make its induction expression in Escherichia coll. [ Result] The target fragment with the length of 2 208 bp was obtained from PCR amplification. The nucleotide homologies between the cloned NS1 gene and the reported relevant PPV genes were from 97.3 % to 99.4 %, which indicated that NS1 gene had high conservation. But it had a 12-basepair successive deletion near the hydroxyl end. The cloned PPV NS1 gene was successfully expressed in prokaryotic cell, and its expression products existed mostly in inclusion bodies. [ Conclusion] The results of SDS-PAGE detection showed that the molecular weight of PPV NS1 protein was 86 KD.
基金Supported by Research Fund of the Doctoral Program of Higher Education (200805720004)Scientific Research Foundation for Returned Scholars, Ministry of Education of China ([2009]1001)~~
文摘[Objective] To construct prokaryotic expression vectors encoding gene Erb3binding protein （EBP1）, which plays important roles in regulating plant organ size from Nervilia fordii （Hance） Schltr. [Methods] PCR products of NfEBP1 with particular restriction sites and expression vectors, pET-28 and pET-16b were digested. Ligation, transformation and selection were performed to construct the recombinant plasmids pET-28-NfEBP1 and pET-16-NfEBP1. The recombinant plasmids were transformed into E. coli BL21 using heat -shock transformation. [Results] Recombinant plasmids pET-28-NfEBP1-1188 and pET-16-NfEBP1-1188 were constructed and transformed into expressional host cells, E. coli BL21, and validated by colony PCR, sequencing and double digestion. [Conclusion] Prokaryotic expression vectors of EBP1 gene from N. fordii were successfully constructed, which laid the foundation for characterization of the gene function.
基金Supported by National Natural Science Foundation of China(30560012)Foundation of General Project of Yunnan Province(2007C261M)Foundation of Yunnan Educational Committee(07C10700)~~
文摘[ Objective ] This study was to express and purify Arabidopsis thaliana heat shock factor HSF1. [ Method ] Using Escherichia coli M15 harboring HSF1 （pQE32/His6-HSF1, pREP4） as experimental materials, HSF1 was induced to express with isopropyl-β-D-galactoside （IPTG） ; then the expression product was purified using Ni-NTA-agarose affinity chromatography and analyzed by SDS-PAGE. [Result] HSF1 of Arabidopsis thaliana was successfully expressed and purified. [ Conclusion] This study provides materials for understanding the blinding site of HSF1 on Arabidopsis thaliana chromosome, further laying a good foundation for revealing the regulatory mechanism and physiological function of HSF1.
文摘Zymography and in situ hybridization were used to investigate matrixmetalloproteinase -2, -9 (MMP -2, MMP-9) activities and expressions of MMP -2, -9 and TIMP1, -2, -3 (tissue inhibitors of matrix metallo-proteinases) mRNA in the rat uterus during estrouscycle. The relative activity was semiquanted by using densitometric analysis. The MMP-2(67 kDa) activity in every stage during estrpus cycle was detected by zymography. MMP-2activity was highest at proestrus; higher at estrus and metaestrus; lowest at diestrus. Throughin situ hybridization, MMP -2, -9, TIMP -1~ -3 mRNA mainly in hasal stroma cells of uterineendometrium were detected. The positive signals of MMP -2 and -9 mRNAs in hasal stromacells were shown stronger at proestrus, estrus and metaestrus while they showed the weakest atdiestrus. The expression of MMP -2 mRNA coincided with MMP -2 activity change. MMP-2and -9 mRNAs were also highly expressed in uterine circular muscle at estrus. Weak signals ofMMP -9 mRNA were detected in uterine luminal and glandular epithelial cells at estrus.TIMP -1 mRNA in hasal stroma cells was shown as the strongest expression at estrus andmetaestrus; stronger at proestrus and the weakest at diestrus. TIMP-2 mRNA in basal stromacells was stronger at estrus and diestrus; weaker at proestrus and metaestrus. TIMP -1 and -2mRNAs were also highly expressed in uterine luminal and glandular epithelial cells at estrus.TIMP -3 mRNA in hasal stroma cells revealed the strongest expression at estrus; stronger atdiestrus and metaestrus and showed the weakest at proestrus. The mRNA was also highlyexpressed in uterine circular muscle at estrus. In short, our present results provide evidencethat MMP -2, -9 and TIMP -1~ -3 were involved in rat uterine endometrium reconstructionduring estrous cycle.
文摘[Objective] The aim of experiment was to lay molecular foundation for studying maturity mechanism of banana after harvest. [Method] The combined method of suppressing subtractive hybridization and cDNA micro-array were used to obtain cDNA segment of one PRMT gene in banana and the whole cDNA sequence of the gene was cloned.The bioinformatics analysis was operated on it,in addition, the expression profile analysis was conducted in different organs and different mature periods of banana.[Result] The whole length of cDNA in MaPRMT1 was 1 158 bp and possessed a complete open reading frame,which could encode 385 amino acids.It had high homology with PRMT in plant,containing one Methyltransf_1 domain.The MaPRMT1 gene was expressed in root,stem,leaf and fruit of banana and the expression levels in stem and leaf were relatively high.As the increase of days after harvest,the expression level declined gradually,however it reached maximum when ethylene release was biggest,then it declined.[Conclusion] MaPRMT1 belonged to the first kind of arginine methyltransferase and it was expressed differently in different organs and fruits at different mature periods.
文摘A genomic DNA fragment containing the 5'-upstream sequence and part of the open reading frame corresponding to the cytosolic fructose-1,6-bisphosphatase (cyFBPase) cDNA was isolated by Genome Walking. The 1 195 lip 5'-flanking region which started from the translation initiation ATG codon was fused to reporter gene encoding beta-glucuronidase (GUS) and stably transferred to rice via particle bombardment. Strong GUS activity was detected in leaves and leaf sheaths of transgenic rice, but not in culms and roots. Histochemical localization revealed that GUS expression was exclusively restricted to mesophyll cells in transgenic rice. Our results indicate that the 1 195 bp fragment contains all the cis-elements required for directing mesophyll-specific expression pattern in rice.
文摘We describe the temporal and spatial expression pattern of Sox 1 gene during Xenopus laevis early development and compare the expression patterns of Sox 1-3 in the developing eye and brain. Alignment of Sox 1-3 amino acid sequences shows a high conservation within the HMG-box DNA binding domains. RT-PCR analysis indicates that Sox 1 is expressed throughout development from the unfertilized egg to at least the tadpole stage, although at different expression levels. The transcripts of XSox 1 are detected in the animal pole at cleavage and blastrula stages and mainly in the central nervous system （CNS） and the developing eye at neurula stages. The study of the developmental expression of XSox 1 will aid in the elucidation of the function of SoxB 1 subgroup genes in vertebrate neurogenesis.
基金Supported by College Student Innovation Fund Project of Jilin University(2015821243)~~
文摘[Objective] This study was conducted to investigate cloning and expression of Pun1 gene controlling pungency of pepper. [Method] With Capsicum annuum L as a material, the cDNA sequence of Capunl gene was obtained, with a total length of 1 457 bp, coding 440 amino acids. [Result] Phylogenetic analysis showed that Capunl was closest to Pun1 of C. chinense, with a genetic distance of 0.019 3. Plant expression vector pCAM-Punl-GFP was constructed and transformed into to- bacco, and it was found that the protein coded by fusion gene Punl：：GFP was lo- cated on cell membrane. Prokaryotic expression vectors were constructed, and by SDS-PAGE and Western Blot detection, an induced protein with a molecular weight of 63 ku was obtained. It was found by real-time fluorescence quantitative expres- sion that Pun1 gene was expressed at the highest level 30 d after flowering, de- creased then, and could not be detected substantially 40 and 45 d after flowering. [Conclusion] This study provides information and reference for molecular regulation mechanism of Pun1 gene.
文摘The Raphanus sativus L. antifungal protein 1 (Rs_AFP1) gene was isolated by polymerase chain reaction (PCR). The complete open reading frame and the fragment encoding the putative mature protein were inserted into the prokaryotic expression vector pET_32b(+), respectively. Subsequent expression showed that the Rs_AFP1 was produced in E. coli as a 27 kD fusion protein only when the N_terminal signal peptide was removed. After treatment with thrombin to remove part of the N_terminal His.tag sequence, the bacterially expressed Rs_AFP1 was used for fungal growth inhibition assay which was conducted on Verticillium dahliae Kleb., a soil_born fungus causing the cotton wilt disease. Results showed that, in the liquid medium, the Rs_AFP1 fusion protein at a concentration of 0.3 g/L clearly inhibited the growth of V. dahliae and the germination of spores. Thus the bacterially expressed fusion protein had the antifungal activity against V. dahliae.
文摘AIM To detect the congenital expression patterns of mdr 1 gene in commonly encountered malignant tumors in clinic, and the relationship between the expression of mdr 1 gene and the prognostic morphology in esophageal carcinomas. METHODS A total of 151 resected samples of malignant tumors without preoperative treatment were taken from Anyang City Tumor Hospital. The congenital expression of their mdr 1 gene was detected with reverse transcription polymerase chain reaction (RT PCR) and was compared with each other. The positive incidence of mdr 1 gene in 46 samples of esophageal carcinoma was compared with their differentiated grades, TNM stages and macroscopic types, and the precautions and advantages of RT PCR were evaluated. RESULTS All the 151 samples were confirmed to be malignant histopathologically, including cancers of stomach and gastric cardia (n =51), esophagus ( n =46), colorectum ( n =16), breast ( n =15), thyroid ( n =10), lung ( n =9), uterine cervix ( n =24). The positive expression rate of their mdr 1 gene was 33 3%, 37%, 31 3%, 13 2%, 40%, 55%, and 0% respectively. All the 46 samples of esophageal carcinoma were pathologically confirmed to be squamous cell carcinoma. The total expression rate of their mdr 1 gene was 37% (17/46), 35% (6/17), 40% (8/20), and 33% (3/9) for differentiation grade Ⅰ, Ⅱ and Ⅲ respectively. The expression rate of TNM classification was 33% (6/18), 40% (5/12) and 37% (6/16) in stage Ⅱa, Ⅱb and Ⅲ. The expression rate was 33% (3/9) in ulcerous type, 37% (3/8) in constrictive one, 33% (5/15) in fungoid one, and 40% (6/14) in medullary one. No statistically significant difference was found. CONCLUSION Compared with other methods, RT PCR is more simple, reliable and accurate in detecting mdr 1 gene expression in tissues of tumor. The overexpression of mdr 1 gene in these neoplasms suggested that cases should be handled differently for chemotherapy with rational use of drugs. Excision is the chief treatment for carcinoma of esophagus. The expression of mdr 1 gene in tissues of esophageal cancer is correlated with the parameters of tumor molecular biology which are independent of histopathological morphology.
