[1]张一折)△,王鹏),张守涛),等.稳定表达HIV1GagPol基因真核表达载体的构建与鉴定*[J].郑州大学学报(医学版),2009,(01):145-147.
 ZHANG Yizhe),WANG Peng),ZHANG Shoutao),et al.Stable expression and identification of GagPol gene from HIV1 in HEK293[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2009,(01):145-147.
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稳定表达HIV1GagPol基因真核表达载体的构建与鉴定*()
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2009年01期
页码:
145-147
栏目:
论著
出版日期:
2009-01-01

文章信息/Info

Title:
Stable expression and identification of GagPol gene from HIV1 in HEK293
作者:
张一折1)王鹏2)张守涛1)刘伟1)刘国祥1)
1)郑州大学生物工程系郑州4500012)吉林省四平市疾病预防控制中心四平136000
Author(s):
ZHANG Yizhe1)WANG Peng2)ZHANG Shoutao1)LIU Wei1)LIU Guoxiang1)
1)Department of Bioengineering,Zhengzhou University,Zhengzhou 4500012)Centers for Disease Control and Prevention and Control,Siping 136000
关键词:
HIV1GagPol基因稳定表达病毒样颗粒疫苗
Keywords:
HIV1 GagPol genestable expressionVLP vaccine
分类号:
R183
摘要:
构建能稳定表达HIV1GagPol基因的真核表达载体。方法:用PCR方法扩增HIV1GagPol基因,克隆入pMD18T,然后亚克隆入真核表达质粒pcDNA3.1(-)中,将构建的重组质粒pcDNA3.1(-)/GagPol分别瞬时和稳定转染HEK293细胞,经G418筛选,建立稳定转染GagPol基因的细胞系,并应用WesternBlot方法鉴定表达产物。结果:酶切鉴定和WesternBlot检测证实重组质粒pcDNA3.1(-)/GagPol构建正确,并能稳定表达HIV1GagPol蛋白。结论:成功构建携带HIV1GagPol基因的真核表达载体pcDNA3.1(-)/GagPol,并在HEK293细胞中获得稳定表达。
Abstract:
To construct a recombinant eukaryotic expression plasmid for stable expression of HIV1 GagPol protein and to detect its expression in HEK293 in order to lay the foundation for further development HIV1 VLP vaccine.Methods:HIV1 GagPol gene was amplified by PCR, then subcloned into eukaryotic expression vector pcDNA3.1(-).The recombinant plasmid pcDNA3.1(-)/GagPol was transfected into HEK293 cell line ,and the expression of the GagPol gene was analyzed by Western Blot.Results: The successful construction of pcDNA3.1(-)/GagPol was verified by restrictive endonuclease assay, and the expression of GagPol gene was detected in the lysate of transfected HEK293 cells by Western Blot.Conclusion: The recombinant plasmid pcDNA3.1(-)/GagPol was successfully constructed and its stable expression in HEK293 cell line was also confirmed in the present study.

参考文献/References:

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备注/Memo

备注/Memo:
*郑州大学人才引进基金资助项目 △男,1975年生,博士,讲师,研究方向:生物工程药物,Email:yizhezhang@126.com
更新日期/Last Update: 2010-05-18