Expression of HaSNPV sod gene in E．coli

GAO Guo—hui; ZHANG Chuan—XI*

PDF
Cite
Share

facebook

twitter

google

linkedin

Volume 18 Issue 5

October 2003

GAO Guo—hui and ZHANG Chuan—XI*. Expression of HaSNPV sod gene in E．coli[J]. Virologica Sinica, 2003, 18(5): 482-485.

Citation: GAO Guo—hui, ZHANG Chuan—XI*. Expression of HaSNPV sod gene in E．coli .VIROLOGICA SINICA, 2003, 18(5) : 482-485.

棉铃虫核型多角体病毒sod基因在大肠杆菌中的表达

高国辉 ,
张传溪

1.
浙江大学应用昆虫学研究所病毒分子生物学实验室，浙江杭州，310029

摘要

摘要：用PCR方法从棉铃虫(Helicoverpa armigera)单粒包埋型核型多角体病毒(HaSNPV) C1株基因组中扩增 sod基因编码区，克隆到pGEM—T-easy vector，测定了核苷酸序列。将基因编码区克隆到原核表达载体pETblue2，构建了含表达质粒pETblue2／HaSNPV SOD，转化大肠杆菌DE3(BL21)进行IPTG诱导表达。SDS—PAGE分析表明SOD 的表达量约为细胞总蛋白的37％。邻苯三酚法测定表达蛋白活性，结果表明每毫克菌体可溶性总蛋白中表达产物校正酶活力单位为694 U／mg。
- HaSNPV：Cu／Zn—SOD
- , 原核表达
- , 活性测定

Expression of HaSNPV sod gene in E．coli

1.
Institute of Applied Entomology,Zhejiang University,Hangzhou 3 1 0029，China

Abstract

Abstract：The coding region of superoxide dismutase was amplified by using PCR method from Helicoverpa armigera single enveloped nucleocapsid nucleopolyhedrovirus(HaSNPV)C 1 genome．The PCR product was cloned into pGEM —T-easy vector and sequenced．Th e cloned coding region of HaSNPV SOD was inserted into the expression vector pET blue一2 to form the recombinant plasmid pETblue2／HaSNPV SOD and was then transformed into E．coli DE3 for expression．The SDS—PAGE an alysis revealed that the expressed recombinan t SOD accumulated up to 37％ of the total bacterial protein．The enzyme activity of HaSNPV SOD was assayed by Pyrogallol autooxidation method．The result showed that the revised enzyme activity of the expressed product in the total soluble protein was 694 U／mg．
- HaSNPV
- , Cu／Zn SOD
- , Prokaryotic expression

References
Proportional views

PDF

Article Metrics

Article views(4062) PDF downloads(851) Cited by(0)

Proportional views

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Expression of HaSNPV sod gene in E．coli

1. Institute of Applied Entomology,Zhejiang University,Hangzhou 3 1 0029，China

Abstract: Abstract：The coding region of superoxide dismutase was amplified by using PCR method from Helicoverpa armigera single enveloped nucleocapsid nucleopolyhedrovirus(HaSNPV)C 1 genome．The PCR product was cloned into pGEM —T-easy vector and sequenced．Th e cloned coding region of HaSNPV SOD was inserted into the expression vector pET blue一2 to form the recombinant plasmid pETblue2／HaSNPV SOD and was then transformed into E．coli DE3 for expression．The SDS—PAGE an alysis revealed that the expressed recombinan t SOD accumulated up to 37％ of the total bacterial protein．The enzyme activity of HaSNPV SOD was assayed by Pyrogallol autooxidation method．The result showed that the revised enzyme activity of the expressed product in the total soluble protein was 694 U／mg．