湖南农业大学学报（自然科学版）2018，Vol.44Issue(2)：157-161,5.DOI:10.13331/j.cnki.jhau.2018.02.008
苎麻香豆酸-3-羟化酶基因的原核表达
Prokaryotic expression of coumarate 3–hydroxylase gene in ramie
摘要
Abstract
The gene BnC3H cloned from ramie and its recombinant proteins expressed in Escherichia coli were analyzed to reveal the enzymatic characteristics and the metabolic mechanism of regulating the lignin in ramie. The open reading frame(ORF)sequence of BnC3H gene was obtained by RT–PCR,and then the cloned genes of BnC3H were inserted into vector pET–22b. The recombinant plasmids pET–22b–C3H were expressed in the prokaryotic expression system after its transformation into E.coli BL21.The result showed that the BnC3H ORF sequence was 1 536 bp and encodes 511 amino acids. The relative molecular weight is 57 800. The optimum concentration of IPTG was 1 mmol/L, results of SDS–PAGE showed that the specific fusion protein was successfully induced by IPTG.关键词
苎麻/苎麻香豆酸–3–羟化酶/基因克隆/原核表达Key words
Boehmeria nivea/boehmeria nivea coumarate 3–hydroxylase/gene cloning/prokaryotic expression分类
生物科学引用本文复制引用
袁有美,郭清泉..苎麻香豆酸-3-羟化酶基因的原核表达[J].湖南农业大学学报（自然科学版）,2018,44(2):157-161,5.基金项目
国家自然科学基金项目(31071457) （31071457）
