新疆农业科学2012，Vol.49Issue(2)：285-289,5.DOI:CNKI:65-1097/S.20120302.2240.016
盐穗木HcGKR基因亚细胞定位表达载体的构建及定位分析
Construction of Expression Vector and Subcellular Localization Analysis of Guanylate Kinase -related Protein Gene in Halostachys caspica
摘要
Abstract
[Objective and Method] In this study green fluorescent protein gene (GFP) was used as reporter gene in biolistic transformation experiment. Subcellular localization expression vector of HcGKR gene was constructed. [Result] Firstly, the HcGKR gene full - length fragment was obtained from pGEM - T - GKR by PCR, then it was cloned into subcellular localization expression vector 35 S: GFP. The subcellular localization expression vector 35 S - GFP - HcGKR is driven by CaMV35S promoter with GFP reporter gene and the target gene, GFP reporter gene and target gene express in single cell. Secondly, verified by digestion with restriction enzymes, the vector 35S - GFP - HcGKR was transformed into onion by gene gun method and the GFP - HcGKR fusion gene expression was detected within 24 h after bombardment under confocal microscopy. [ Conclusion ] The result of subcellular localization expression showed that the gene was located in cell membrane. The study of this paper has laid a strong foundation for analyzing function of HcGKR gene in Halostachys caspica in the future.关键词
盐穗木/GKR基因/绿色荧光蛋白(GFP)/载体构建/亚细胞定位Key words
Halostachys caspica/ guanylate kinase - related gene/ green fluorescent protein ( GFP) /vector construction/ subcellular localization分类
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郝晓燕,张毓露,足木热木,危晓薇,邵琳,陈果,黄全生..盐穗木HcGKR基因亚细胞定位表达载体的构建及定位分析[J].新疆农业科学,2012,49(2):285-289,5.基金项目
新疆维吾尔自治区“十一五”重大专项子项(200731138-3) （200731138-3）
新疆维吾尔自治区农作物生物技术重点实验室开放课题(XJDX0201 -2011 -05) （XJDX0201 -2011 -05）
