日本血吸虫谷胱甘肽硫转移酶原核表达载体的构建及序列分析  

Molecular Cloning and Sequence Analysis of Glutathione S-transferase Gene in Schistosoma japonicum

作　　者：方桂杰[1] 乔宪凤[2] 

机构地区：[1]湖北工业大学生物工程学院发酵工程省部共建教育部重点实验室,湖北武汉430068 [2]湖北省农业科学院畜牧兽医研究所动物胚胎工程与分子育种湖北省重点实验室,湖北武汉430064

出　　处：《江西农业学报》2010年第11期8-10,14,共4页Acta Agriculturae Jiangxi

基　　金：湖北工业大学博士启动基金(32700224)

摘　　要：为构建带有组氨酸标签(his-tag)的日本血吸虫谷胱甘肽硫转移酶(sjGST)表达载体。以载有sjGST基因的载体PGEX-KG为模板,PCR扩增sjGST基因,并在进行酶切和基因测序后将其克隆到pET28a载体上。运用生物学软件与具有代表性的血吸虫GST基因进行比对并对可能的表达产物进行分析。结果表明,PCR产物与3种sjGST基因有99%的同源性。重组载体构建成功。In order to construct the recombinant clone of glutathione S-transferase gene(GST) in Schistosoma japonicum and express the recombinant sjGST protein with his-tag that could be purified by single-step Ni-NTA affinity chromatography,the DNA fragment of sjGST gene was amplified by PCR with template PGEX-KG.Then it was cloned into vector pET28a,and the recombinant prokaryotic expressing vector was constructed and corroborated through restriction enzymes map and sequencing.Identity analysis and the primary and tertiary structure of translation alignment were made by using NCBI-blast and Swiss-module.Results showed that pET28a-sjGST recombinant clone was constructed successfully.The length of amplified GST gene has the opening reading frame(ORF) of 687 bp in length and has 99% identity with PGEX-KG-GST,S.japonicum-GST-Mainland-26ku(sjGST-ML-26) and S.japonicum-GST-Philippines-26ku(sjGST-PL-26).

关 键 词：日本血吸虫 谷胱甘肽硫转移酶 原核 表达载体的构建 序列分析 SCHISTOSOMA JAPONICUM Sequence Analysis 基因测序 重组载体构建 组氨酸标签 生物学软件 表达产物 PCR产物 HIS-TAG GST基因 同源性 代表性 pET28a 模板 酶切 

分 类 号：S[农业科学]