Prokaryotic cloning and expression of HPV16 E7 gene

JIANG Hai-rong; et al

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Prokaryotic cloning and expression of HPV16 E7 gene

更新时间：2024-01-17

- Prokaryotic cloning and expression of HPV16 E7 gene
- Journal of Chongqing Medical University Vol. 34, Issue 9, Pages: 1215-1217(2009)
- 作者机构：
  
  1. 重庆理工大学化学与生物工程学院
  2. 重庆市江北区第一人民医院检验科
  3. 重庆医科大学图书馆文献检索教研室
  4. 重庆市第四人民医院妇产科
  5. 重庆市第四人民医院麻醉科
- 作者简介：
- 基金信息：
- DOI：
  CLC： R346
- Published：2009，
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[1]姜海蓉,彭方毅,刘芳,杜志银,林治华,彭方亮,赵卫兵.HPV16E7基因的原核克隆及表达[J].重庆医科大学学报,2009,34(09):1215-1217.

JIANG Hai-rong, et al. Prokaryotic cloning and expression of HPV16 E7 gene. [J]. Journal of Chongqing Medical University 34(9):1215-1217(2009)
[1]姜海蓉,彭方毅,刘芳,杜志银,林治华,彭方亮,赵卫兵.HPV16E7基因的原核克隆及表达[J].重庆医科大学学报,2009,34(09):1215-1217. DOI：

JIANG Hai-rong, et al. Prokaryotic cloning and expression of HPV16 E7 gene. [J]. Journal of Chongqing Medical University 34(9):1215-1217(2009) DOI：

摘要

目的:探讨从喉癌组织中HPV16E7蛋白的原核克隆及表达。方法:采用PCR方法扩增HPV16E7基因

经BamHⅠ和HindⅢ双酶切后插入相同酶切pET28a（+）载体

转化JM109感受态细胞

并进行阳性克隆筛选。经IPTG对重组质粒进行蛋白诱导表达

SDS-PAGE和Western blot检测目标蛋白表达情况。结果:成功构建了原核表达质粒pET28/E7、HPV16E7融合蛋白在BL21（DE3）菌株中高效表达。结论:HPV16E7融合蛋白的表达为进一步深入研究HPV16E7蛋白的生物学特性及其致细胞转化的作用机制奠定了基础。

Abstract

Objective:To explore the cloning and expression of HPV16 E7 protein from laryngeal carcinoma. Methods: HPV16 E7 gene was amplified by PCR. The amplified fragment was inserted into the plasmid pET28a (+) digested with BamHⅠand Hind Ⅲ. The recombinant plasmid pET28/E7 was transformed into E.coli JM109 which was selected with ampicillin. HPV16 E7 recombinant protein expression in the E.coli BL21(DE3) was identified by SDS-PAGE and Western blot. Results: The prokaryotic recombinant plasmid pET28/E7 was successfully constructed. The BL21(DE3) transformed recombinant plasmid pET28/E7 had expressed HPV16 E7 recombinant protein effectively. Conclusion:The construction of the prokaryotic recombinant plasmid pET28/E7 and the successful expression of the recombinant protein HPV16 E7 pave way for the profound research of the biological properties and the transformational mechanism of the HPV16 E7 protein on the specific cells.

关键词

HPV16E7原核表达SDS-PAGEWestern blot

Keywords

HPV16-E7prokaryotic expressionSDS-PAGEWestern blot

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