Construction and expression of secreted chimericAFP-HSP70 plasmid in eukaryotic cells

LAN Yinghua; et al

doi:10.13406/j.cnki.cyxb.2004.06.008

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Construction and expression of secreted chimericAFP-HSP70 plasmid in eukaryotic cells

更新时间：2024-01-17

- Construction and expression of secreted chimericAFP-HSP70 plasmid in eukaryotic cells
- Journal of Chongqing Medical University Issue 6, Pages: 734-737(2004)
- 作者机构：
  
  1. 重庆医科大学病毒性肝炎研究所
  2. 重庆医科大学病毒性肝炎研究所,重庆,400010
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2004.06.008
  CLC： R73-3
- Published：2004，
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[1]兰英华,唐俐,陈敏,彭明利,任红.甲胎蛋白与热休克蛋白70基因融合载体的构建及表达[J].重庆医科大学学报,2004(06):734-737+741.

LAN Yinghua, et al. Construction and expression of secreted chimericAFP-HSP70 plasmid in eukaryotic cells. [J]. Journal of Chongqing Medical University (6):734-737(2004)
[1]兰英华,唐俐,陈敏,彭明利,任红.甲胎蛋白与热休克蛋白70基因融合载体的构建及表达[J].重庆医科大学学报,2004(06):734-737+741. DOI： 10.13406/j.cnki.cyxb.2004.06.008.

LAN Yinghua, et al. Construction and expression of secreted chimericAFP-HSP70 plasmid in eukaryotic cells. [J]. Journal of Chongqing Medical University (6):734-737(2004) DOI： 10.13406/j.cnki.cyxb.2004.06.008.

摘要

目的 :构建分泌性小鼠甲胎蛋白（α -Fetoprotein

AFP）与结核杆菌热休克蛋白 70 （mycobacteriumtuberculosisheatshockprotein 70

Mt.HSP70 ）基因融合载体

并研究其在真核细胞中的表达情况。方法 :以 pSecTag2 B为基本单位构建AFP及与HSP70的融合表达载体

融合基因以G -S -G -G -S连接子连接 ;用脂质体将系列载体导入COS - 7细胞

4 8h后以免疫化学方法检测其表达

抽提总RNA作RT -PCR测其在转录水平的表达。结果 :构建的AFP和HSP70的单独及融合表达载体导入COS - 7细胞 4 8h后经细胞免疫化学染色为阳性

RT -PCR可扩增出特异性条带。结论 :AFP和HSP70的单独及融合表达载体构建成功

能在真核细胞中表达。

Abstract

Objective:To construct secreted chimeric vector of AFP-HSP70 and investigate transient expression of vectors in eukaryotic cells.Methods:Vectors were constructed by routine molecular technique.Fusion genes were linked with G-S-G-G-S linker.Vectors were transfected into COS-7 cells by Lipofectamine 2000. Proteins were assayed by immunocytochemistry and RT-PCR for its RNA.Results:Proteins expressed in COS-7 were comfirmed by immunocytochemistry 48 hours later and RNA was detected by RT-PCR.Conclusion:AFP-HSP70 chimeric vector is constructed successfully and could be expressed in eukaryotic cells.

关键词

甲胎蛋白热休克蛋白70癌肝细胞融合

Keywords

FetoproteinHeat shock protein 70HepatocellularcarcinomaFusion

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