Abstract The dopamine transporter （DAT） is involved in the regulation of extracellular dopamine levels. A 40-bp variable-number tandem repeat （VNTR） polymorphism in the 3-untranslated region （3UTR） of the DAT ha...Abstract The dopamine transporter （DAT） is involved in the regulation of extracellular dopamine levels. A 40-bp variable-number tandem repeat （VNTR） polymorphism in the 3-untranslated region （3UTR） of the DAT has been reported to be associated with various phenotypes that are involved in the aberrant regulation of dopaminergic neu- rotransmission. In the present study, we found that miR- 137 and miR-491 caused a marked reduction of DAT expression, thereby influencing neuronal dopamine trans- port. Moreover, the regulation of miR-137 and miR-491 on this transport disappeared after the DAT was silenced. The miR-491 seed region that is located on the VNTR sequence in the 3＇UTR of the DAT and the regulatory effect of miR- 491 on the DAT depended on the VNTR copy-number. These data indicate that miR-137 and miR-491 regulate DAT expression and dopamine transport at the post- transcriptional level, suggesting that microRNA may be targeted for the treatment of diseases associated with DAT dysfunction.展开更多
The eukaryotic vectors VR1012 expressing survivin or 33 tandem repeats of human mucin 1（MUC1）（VNTRs）,namely,VR1012-S and VR1012-VNTR（VNTR=variable number of tandem repeat）,were constructed by cloning survivin an...The eukaryotic vectors VR1012 expressing survivin or 33 tandem repeats of human mucin 1（MUC1）（VNTRs）,namely,VR1012-S and VR1012-VNTR（VNTR=variable number of tandem repeat）,were constructed by cloning survivin and VNTR genes into VR1012,respectively.The eukaryotic vector pEGFP expressing survivin and MUC1 VNTRs fusion gene pEGFP-MS was also constructed.Mouse melanoma cell line（B16） stably expressing survivin and MUC1 VNTRs（MS ＋ B16） was established by Lipofectamine-mediated transfection of pEGFP-MS into B16 cells.EGFP expression in MS ＋ B16 cells was observed using a fluorescent microscope and survivin and MUC1 VNTRs（MS） expression was confirmed by means of Western blot analysis.A syngenic graft tumor model was generated by subcutaneous injection of MS ＋ B16 cells into C57/BL6 mice and tumor size increased rapidly with time in a cell number dependent manner.After the third immunization,mice were challenged subcutaneously with 5×l0 5 MS ＋ B16 cells.Compared with that of the negative control immunized with phosphate-buffered saline（PBS）,a significant reduction of tumor growth was observed in groups immunized with survivin plasmid DNA and MUC1 VNTRs plasmid DNA.Thus,the suppression of subcutaneous tumor was antigen-specific.This model is useful for the development of tumor vaccines targeting survivin and MUCI VNTRs.展开更多
Background:The prevalence of human brucellosis in Qinghai Province of China has been increasing rapidly,with confirmed cases distributed across 31 counties.However,the epidemiology of brucellosis transmission has not ...Background:The prevalence of human brucellosis in Qinghai Province of China has been increasing rapidly,with confirmed cases distributed across 31 counties.However,the epidemiology of brucellosis transmission has not been fully elucidated.To characterize the infecting strains isolated from humans,multiple-locus variable-number tandem repeats analysis(MLVA)and whole-genome single-nucleotide polymorphism(SNP)-based approaches were employed.Methods:Strains were isolated from two males blood cultures that were confirmed Brucella m elitensis positive following biotyping and MLVA.Genomic DNA was extracted from these two strains,and whole-genome sequencing was performed.Next,SNP-based phylogenetic analysis was performed to compare the two strains to 94 B.melitensis strains(complete genome and draft genome)retrieved from online databases.Results:The two Brucella isolates were identified as B.melitensis biovar 3(QH2019001 and QH2019005)following conventional biotyping and were found to have differences in their variable number tandem repeats(VNTRs)using MLVA-16.Phylogenetic examination assigned the 96 strains to five genotype groups,with QH2019001 and QH2019005 assigned to the same group,but different subgroups.Moreover,the QH2019005 strain was assigned to a new subgenotype,llj,within genotype II.These findings were then combined to determine the geographic origin of the two Brucella strains.Conclusions:Utilizing a whole-genome SNP-based approach enabled differences between the two B.m elitensis strains to be more clearly resolved,and facilitated the elucidation of their different evolutionary histories.This approach also revealed that QH2019005 is a member of a new subgenotype(Hj)with an ancient origin in the eastern Mediterranean Sea.展开更多
基金supported by grants from the National Postdoctoral Science Foundation,China(2014M552219)the Natural Science Foundation of Guangdong Province,China(2015 A030313889,2015A030401013,2014A030313709,and 2014A030 313710)+1 种基金the Science and Technology Planning Project of Shenzhen Municipality,China(ZDSYS201504301045406,JCYJ20150403110 829621,JCYJ20150403091443301,JCYJ20140415162542975,JCYJ 20140415162338855,JCYJ20140828163634004,and JCYJ201206 16144352139)the Health and Family Planning Commission Project of Shenzhen Municipality,China(201401026)
文摘Abstract The dopamine transporter （DAT） is involved in the regulation of extracellular dopamine levels. A 40-bp variable-number tandem repeat （VNTR） polymorphism in the 3-untranslated region （3UTR） of the DAT has been reported to be associated with various phenotypes that are involved in the aberrant regulation of dopaminergic neu- rotransmission. In the present study, we found that miR- 137 and miR-491 caused a marked reduction of DAT expression, thereby influencing neuronal dopamine trans- port. Moreover, the regulation of miR-137 and miR-491 on this transport disappeared after the DAT was silenced. The miR-491 seed region that is located on the VNTR sequence in the 3＇UTR of the DAT and the regulatory effect of miR- 491 on the DAT depended on the VNTR copy-number. These data indicate that miR-137 and miR-491 regulate DAT expression and dopamine transport at the post- transcriptional level, suggesting that microRNA may be targeted for the treatment of diseases associated with DAT dysfunction.
基金Supported by the National Natural Science Foundation of China(No.30872396)the Scientific Research Foundation of Jilin Province,China(Nos.20080709,200905169)the Jilin University Basic Research Project,China(No.200903255)
文摘The eukaryotic vectors VR1012 expressing survivin or 33 tandem repeats of human mucin 1（MUC1）（VNTRs）,namely,VR1012-S and VR1012-VNTR（VNTR=variable number of tandem repeat）,were constructed by cloning survivin and VNTR genes into VR1012,respectively.The eukaryotic vector pEGFP expressing survivin and MUC1 VNTRs fusion gene pEGFP-MS was also constructed.Mouse melanoma cell line（B16） stably expressing survivin and MUC1 VNTRs（MS ＋ B16） was established by Lipofectamine-mediated transfection of pEGFP-MS into B16 cells.EGFP expression in MS ＋ B16 cells was observed using a fluorescent microscope and survivin and MUC1 VNTRs（MS） expression was confirmed by means of Western blot analysis.A syngenic graft tumor model was generated by subcutaneous injection of MS ＋ B16 cells into C57/BL6 mice and tumor size increased rapidly with time in a cell number dependent manner.After the third immunization,mice were challenged subcutaneously with 5×l0 5 MS ＋ B16 cells.Compared with that of the negative control immunized with phosphate-buffered saline（PBS）,a significant reduction of tumor growth was observed in groups immunized with survivin plasmid DNA and MUC1 VNTRs plasmid DNA.Thus,the suppression of subcutaneous tumor was antigen-specific.This model is useful for the development of tumor vaccines targeting survivin and MUCI VNTRs.
基金supported by National Key R&D Program of China(Grant No.2020YFA0907101)Major Infectious Diseases such as AIDS and Viral Hepatitis Prevention and Control Technology Major Projects(2018ZX10201002)the National Natural Scientific Foundation of China(81860588).
文摘Background:The prevalence of human brucellosis in Qinghai Province of China has been increasing rapidly,with confirmed cases distributed across 31 counties.However,the epidemiology of brucellosis transmission has not been fully elucidated.To characterize the infecting strains isolated from humans,multiple-locus variable-number tandem repeats analysis(MLVA)and whole-genome single-nucleotide polymorphism(SNP)-based approaches were employed.Methods:Strains were isolated from two males blood cultures that were confirmed Brucella m elitensis positive following biotyping and MLVA.Genomic DNA was extracted from these two strains,and whole-genome sequencing was performed.Next,SNP-based phylogenetic analysis was performed to compare the two strains to 94 B.melitensis strains(complete genome and draft genome)retrieved from online databases.Results:The two Brucella isolates were identified as B.melitensis biovar 3(QH2019001 and QH2019005)following conventional biotyping and were found to have differences in their variable number tandem repeats(VNTRs)using MLVA-16.Phylogenetic examination assigned the 96 strains to five genotype groups,with QH2019001 and QH2019005 assigned to the same group,but different subgroups.Moreover,the QH2019005 strain was assigned to a new subgenotype,llj,within genotype II.These findings were then combined to determine the geographic origin of the two Brucella strains.Conclusions:Utilizing a whole-genome SNP-based approach enabled differences between the two B.m elitensis strains to be more clearly resolved,and facilitated the elucidation of their different evolutionary histories.This approach also revealed that QH2019005 is a member of a new subgenotype(Hj)with an ancient origin in the eastern Mediterranean Sea.
