[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
The use of pan-ethnic-group products form knowledge primarily depends on a designer＇s subjective experience without user participation. The majority of studies primarily focus on the detection of the perceptual deman...The use of pan-ethnic-group products form knowledge primarily depends on a designer＇s subjective experience without user participation. The majority of studies primarily focus on the detection of the perceptual demands of consumers from the target product category. A pan-ethnic-group products form gene clustering method based on emotional semantic is constructed. Consumers＇ perceptual images of the pan-ethnic-group products are obtained by means of product form gene extraction and coding and computer aided product form clustering technology. A case of form gene clustering about the typical pan-ethnic-group products is investigated which indicates that the method is feasible. This paper opens up a new direction for the future development of product form design which improves the agility of product design process in the era of Industry 4.0.展开更多
Dear Editor,Understanding the complex interactions between transcription factors(TFs)and DNA is crucial in various scientific fields,including genetics,molecular biology,and biochemistry.Current methodologies such as ...Dear Editor,Understanding the complex interactions between transcription factors(TFs)and DNA is crucial in various scientific fields,including genetics,molecular biology,and biochemistry.Current methodologies such as yeast one-hybrid,chromatin immunoprecipitation followed by sequencing,the firefly luciferase(Luc)reporter system,and DNA electrophoretic mobility shift assays often necessitate complicated procedures and costly chemicals(Ferraz et al.,2021).The adoption of the pigmentation-based reporter RUBY has facilitated the study of various plant signaling processes due to the easily observable nature of pigments(He et al.,2020).However,pigmentation-based assays lack sensitivity,and their high background limits broader applications.The recent discovery of the fungal Luc coding gene Luz in the bioluminescence pathway from the fungus Neonothopanus nambi offers new opportunities for enhancing reporter systems in transcriptional regulation assays(Kotlobay et al.,2018).The fungal bioluminescence pathway(FBP)is a cyclic metabolic route that involves the action of four enzymes:HispS,H3H,Luz,and CPH(Figure 1A).The pathway’s initial committed step utilizes caffeic acid as a substrate by the fungus HispS;NPGA from Aspergillus nidulans is capable of post-translationally modifying HispS.This modification enhances the activity of HispS,thereby increasing the intensity of FBP(Khakhar et al.,2020;Zheng et al.,2023;Shakhova et al.,2024).Notably,the synthesis pathway of caffeic acid has evolutionarily conserved homologs in plants(Supplemental Figure 1),suggesting that plant cells might independently utilize their own metabolic pool of caffeic acid to support the FBP without the need for additional substrate.This finding underscores the potential of FBP as a promising reporter for plant transcriptional regulation without substrate addition.展开更多
Noncoding“junk DNA”,which constitutes 98%of our genome,is now generally considered to play a fundamental role in the precise regulation of coding genes to establish cell identity.One feature of functional“junk DNA...Noncoding“junk DNA”,which constitutes 98%of our genome,is now generally considered to play a fundamental role in the precise regulation of coding genes to establish cell identity.One feature of functional“junk DNA”is its accessibility.In cancer cells,aberrant chromatin accessibility is recognized as one of the major hallmarks.Understanding such events requires high-throughput screening,such as Assay for Transposase-Accessible Chromatin using sequencing(ATAC-seq),which generates large-scale data that are puzzling for biologists.However,existing web tools only support cell line data and lack high-quality clinical phenotypes or matched transcriptomes.Here,we developed Shiny Pan-cancer Accessible Chromatin Explorer(SPACE)as an all-in-one web server encompassing 562,709 regulatory elements in 404 patients across 23 cancer types.展开更多
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 Key Technology R&D Program,China(Grant No.2015BAH21F01)National 111 Project,China(Grant No.B13044)
文摘The use of pan-ethnic-group products form knowledge primarily depends on a designer＇s subjective experience without user participation. The majority of studies primarily focus on the detection of the perceptual demands of consumers from the target product category. A pan-ethnic-group products form gene clustering method based on emotional semantic is constructed. Consumers＇ perceptual images of the pan-ethnic-group products are obtained by means of product form gene extraction and coding and computer aided product form clustering technology. A case of form gene clustering about the typical pan-ethnic-group products is investigated which indicates that the method is feasible. This paper opens up a new direction for the future development of product form design which improves the agility of product design process in the era of Industry 4.0.
基金financially supported by the National Natural Science Foundation of China(32470288)the Zhejiang University Global Partnership Fund.
文摘Dear Editor,Understanding the complex interactions between transcription factors(TFs)and DNA is crucial in various scientific fields,including genetics,molecular biology,and biochemistry.Current methodologies such as yeast one-hybrid,chromatin immunoprecipitation followed by sequencing,the firefly luciferase(Luc)reporter system,and DNA electrophoretic mobility shift assays often necessitate complicated procedures and costly chemicals(Ferraz et al.,2021).The adoption of the pigmentation-based reporter RUBY has facilitated the study of various plant signaling processes due to the easily observable nature of pigments(He et al.,2020).However,pigmentation-based assays lack sensitivity,and their high background limits broader applications.The recent discovery of the fungal Luc coding gene Luz in the bioluminescence pathway from the fungus Neonothopanus nambi offers new opportunities for enhancing reporter systems in transcriptional regulation assays(Kotlobay et al.,2018).The fungal bioluminescence pathway(FBP)is a cyclic metabolic route that involves the action of four enzymes:HispS,H3H,Luz,and CPH(Figure 1A).The pathway’s initial committed step utilizes caffeic acid as a substrate by the fungus HispS;NPGA from Aspergillus nidulans is capable of post-translationally modifying HispS.This modification enhances the activity of HispS,thereby increasing the intensity of FBP(Khakhar et al.,2020;Zheng et al.,2023;Shakhova et al.,2024).Notably,the synthesis pathway of caffeic acid has evolutionarily conserved homologs in plants(Supplemental Figure 1),suggesting that plant cells might independently utilize their own metabolic pool of caffeic acid to support the FBP without the need for additional substrate.This finding underscores the potential of FBP as a promising reporter for plant transcriptional regulation without substrate addition.
基金supported by the National Natural Science Foundation of China(31770935,81873531,31970616)the Distinguished Professorship Program of Jiangsu Province to YF+2 种基金the Distinguished Professorship Program of Jiangsu Province to RMthe National Undergraduate Training Programs for Innovation(201710304030Z)the National Undergraduate Training Programs for Innovation(201810304026Z).
文摘Noncoding“junk DNA”,which constitutes 98%of our genome,is now generally considered to play a fundamental role in the precise regulation of coding genes to establish cell identity.One feature of functional“junk DNA”is its accessibility.In cancer cells,aberrant chromatin accessibility is recognized as one of the major hallmarks.Understanding such events requires high-throughput screening,such as Assay for Transposase-Accessible Chromatin using sequencing(ATAC-seq),which generates large-scale data that are puzzling for biologists.However,existing web tools only support cell line data and lack high-quality clinical phenotypes or matched transcriptomes.Here,we developed Shiny Pan-cancer Accessible Chromatin Explorer(SPACE)as an all-in-one web server encompassing 562,709 regulatory elements in 404 patients across 23 cancer types.
