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Position: Home > Articles > Construction of prokaryotic expression vector containing tobacco 4CL and Polygonum cuspidatum STSfusion gene Journal of Beijing University of Agriculture 2013,28 (3) 1-5

烟草4CL与虎杖STS融合基因原核表达载体的构建

作  者:
张红;刘霄霄;解语晨;马雅迪;杨静;马兰青;杨明峰;王有年
单  位:
北京农学院植物科学技术学院农业部都市农业北方重点实验室
关键词:
次生代谢;白藜芦醇;引物悬挂PCR;融合基因;双功能酶
摘  要:
植物次生代谢产物白藜芦醇具有抗肿瘤、保护心血管、抗氧化的药理作用。通过分子生物学技术利用带有13个氨基酸的链接序列(linker)将来自烟草的4-香豆酰辅酶A连接酶(4-Coumarate-CoA ligase,4CL)和虎杖的白藜芦醇合酶(Stilbene synthase,STS)的基因连接,构建成融合基因,并转入原核表达载体。为后续的白藜芦醇的代谢工程奠定了基础。但该研究中所构建的融合基因与原基因相比有7个碱基突变,导致4个氨基酸的改变。这些突变是否会影响到融合酶的活性,试验所引入的13个氨基酸的linker对于融合蛋白活性的影响等都还待后续进一步研究。
译  名:
Construction of prokaryotic expression vector containing tobacco 4CL and Polygonum cuspidatum STSfusion gene
作  者:
ZHANG Hong,LIU Xiaoxiao,XIE Yuchen,MA Yadi,YANG Jing,MA Lanqing,YANG Mingfeng,WANG Younian(Beijing University of Agricultural College of Plant Science and Technology,Key Laboratory of Urban Agriculture(North)Ministry of Agriculture,Beijing 102206,China)
关键词:
plant secondary metabolites;resveratrol;overlap PCR;fusion gene;bifunctional enzyme
摘  要:
The pharmacological function of resveratrol is anti-tumor,cardiovascular protection,antioxidant.In this paper,the fusion gene Nt4CL::PcSTS was constucted by using a link sequence(linker)of 13amino acids.The gene was cloned into the prokaryotic expression vector pET30a.But the sequencing results showed that there were 7nucleotide mutation sites in the fusion gene,result in four amino acid changes.Further experiment will be interesting to the changes and linker which affect fusion protein activity.

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