Objective To construct the zinc finger protein-activating transcription factor (ZFP-ATF) plasmid and evaluate its efficacy in inducing vascular endothelial growth factor (VEGF) expression in EY.HY926 endothelial cells...Objective To construct the zinc finger protein-activating transcription factor (ZFP-ATF) plasmid and evaluate its efficacy in inducing vascular endothelial growth factor (VEGF) expression in EY.HY926 endothelial cells. Methods Firstly, we constructed the ZFP-ATF plasmid, then testified the quantity of VEGF protein in EY.HY926 endothelial cells after transfected with ZFP-ATP plasmid by Western blot, finally, we used the RT-PCR to testify whether the ZFP-ATF can stimulate expression of VEGF splice variants. Results The ZFP-ATF DNA sequences were located the multiclone sites of PVAX1 vector between the site of BamH Ⅰ and Xhol. Western blot result showed VEGF expression in EY.HY926 endothelial cells transfected with ZFP-ATF plasmid was significantly higher than that in cells transfected with VEGF165 (19.95±3.95 vs. 12.15±1.55 μg/μL, P<0.01). RT-PCR result showed VEGF-A mRNA expression level induced by ZFP-ATF was high than that induced by VEGF165. Conclusion ZFP-ATF can up-regulate the VEGF-A expression in comparison with VEGF165, which might have beneficial effects in angiogenesis process.展开更多
The precise roles of the B-box zinc finger family of transcription factors in plant stress are poorly understood.Functional analysis was performed on AtCOL4,an Arabidopsis thaliana L.CONSTANS-like 4 protein that is a ...The precise roles of the B-box zinc finger family of transcription factors in plant stress are poorly understood.Functional analysis was performed on AtCOL4,an Arabidopsis thaliana L.CONSTANS-like 4 protein that is a putative novel transcription factor,and which contains a predicted transcriptional activation domain.Analyses of an AtCOL4 promoter-b-glucuronidase（GUS） construct revealed substantial GUS activity in whole seedlings.The expression of AtCOL4 was strongly induced by abscisic acid（ABA）,salt,and osmotic stress.Mutation in atcol4 resulted in increased sensitivity to ABA and salt stress during seed germination and the cotyledon greening process.In contrast,AtCOL4-overexpressing plants were less sensitive to ABA and salt stress compared to the wild type.Interestingly,in the presence of ABA or salt stress,the transcript levels of other ABA biosynthesis and stress-related genes were enhanced induction in AtCOL4-overexpressing and WT plants,rather than in the atcol4 mutant.Thus,AtCOL4 is involved in ABA and salt stress response through the ABA-dependent signaling pathway.Taken together,these findings provide compelling evidence that AtCOL4 is an important regulator for plant tolerance to abiotic stress.e展开更多
基金Supported by the National Natural Science Foundation of China(81270399and81100226)
文摘Objective To construct the zinc finger protein-activating transcription factor (ZFP-ATF) plasmid and evaluate its efficacy in inducing vascular endothelial growth factor (VEGF) expression in EY.HY926 endothelial cells. Methods Firstly, we constructed the ZFP-ATF plasmid, then testified the quantity of VEGF protein in EY.HY926 endothelial cells after transfected with ZFP-ATP plasmid by Western blot, finally, we used the RT-PCR to testify whether the ZFP-ATF can stimulate expression of VEGF splice variants. Results The ZFP-ATF DNA sequences were located the multiclone sites of PVAX1 vector between the site of BamH Ⅰ and Xhol. Western blot result showed VEGF expression in EY.HY926 endothelial cells transfected with ZFP-ATF plasmid was significantly higher than that in cells transfected with VEGF165 (19.95±3.95 vs. 12.15±1.55 μg/μL, P<0.01). RT-PCR result showed VEGF-A mRNA expression level induced by ZFP-ATF was high than that induced by VEGF165. Conclusion ZFP-ATF can up-regulate the VEGF-A expression in comparison with VEGF165, which might have beneficial effects in angiogenesis process.
基金supported in part by a grant to C.S.K.from the Next-Generation Bio Green21 program (SSAC,PJ00949104)funded by the Rural Development Administration+1 种基金Basic Science Research Programfunded by the Ministry of Education,Science and Technology of Korea (NRF-2010-0022026)
文摘The precise roles of the B-box zinc finger family of transcription factors in plant stress are poorly understood.Functional analysis was performed on AtCOL4,an Arabidopsis thaliana L.CONSTANS-like 4 protein that is a putative novel transcription factor,and which contains a predicted transcriptional activation domain.Analyses of an AtCOL4 promoter-b-glucuronidase（GUS） construct revealed substantial GUS activity in whole seedlings.The expression of AtCOL4 was strongly induced by abscisic acid（ABA）,salt,and osmotic stress.Mutation in atcol4 resulted in increased sensitivity to ABA and salt stress during seed germination and the cotyledon greening process.In contrast,AtCOL4-overexpressing plants were less sensitive to ABA and salt stress compared to the wild type.Interestingly,in the presence of ABA or salt stress,the transcript levels of other ABA biosynthesis and stress-related genes were enhanced induction in AtCOL4-overexpressing and WT plants,rather than in the atcol4 mutant.Thus,AtCOL4 is involved in ABA and salt stress response through the ABA-dependent signaling pathway.Taken together,these findings provide compelling evidence that AtCOL4 is an important regulator for plant tolerance to abiotic stress.e
