Construction of the eukaryotic expression vector of proliferin related protein and preliminary study on its biological activity in 293FT cells

WANG Qi; et al

doi:10.13406/j.cnki.cyxb.2011.04.010

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Construction of the eukaryotic expression vector of proliferin related protein and preliminary study on its biological activity in 293FT cells

更新时间：2024-01-17

- Construction of the eukaryotic expression vector of proliferin related protein and preliminary study on its biological activity in 293FT cells
- Journal of Chongqing Medical University Vol. 36, Issue 4, Pages: 444-446(2011)
- 作者机构：
  
  1. 重庆医科大学公共卫生学院卫生毒理学教研室
  2. 重庆医科大学附属第一医院实验研究中心
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2011.04.010
  CLC： R341
- Published：2011，
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[1]王琦,郝杰,赵丽娜,吕自兰,王丽敏,余秋波,王应雄,黎刚.PRP基因绿色荧光蛋白载体构建及其生物学功能初步研究[J].重庆医科大学学报,2011,36(04):444-446.

WANG Qi, et al. Construction of the eukaryotic expression vector of proliferin related protein and preliminary study on its biological activity in 293FT cells. [J]. Journal of Chongqing Medical University 36(4):444-446(2011)
[1]王琦,郝杰,赵丽娜,吕自兰,王丽敏,余秋波,王应雄,黎刚.PRP基因绿色荧光蛋白载体构建及其生物学功能初步研究[J].重庆医科大学学报,2011,36(04):444-446. DOI： 10.13406/j.cnki.cyxb.2011.04.010.

WANG Qi, et al. Construction of the eukaryotic expression vector of proliferin related protein and preliminary study on its biological activity in 293FT cells. [J]. Journal of Chongqing Medical University 36(4):444-446(2011) DOI： 10.13406/j.cnki.cyxb.2011.04.010.

摘要

目的:构建Prolifein related protein（PRP）基因荧光表达载体

并初步探讨其生物学功能。方法:提取小鼠睾丸RNA

RT-PCR扩增PRP基因编码区片段

酶切连入pEGFP-C1载体

构建含有PRP基因片段的绿色荧光表达载体PRP-pEGFP-C1。为了探讨PRP基因功能

PRP-pEGFP-C1经Lipofectamine 2000介导转染293FT细胞

MTT实验分析细胞增殖变化

流式细胞仪分析细胞周期分布。结果:质粒PRP-pEGFP-C1经测序验证

证实PRP基因已正确连入pEGFP-C1载体。免疫荧光染色显示PRP定位于293FT细胞胞浆。MTT结果表明

上调PRP基因引起293FT细胞增殖活性下降

细胞周期分布情况提示G1期细胞增加

S期细胞减少。结论:成功构建PRP-pEGFP-C1荧光表达载体

对其功能研究表明PRP具有抗人293FT细胞增殖

扰乱细胞增殖周期的功能。本研究为PRP在抗人肿瘤方面的研究提供基础。

Abstract

Objective:To construct the eukaryotie expression vector of proliferin related protein

and to investigate its biological function.Methods:The coding region of proliferin related protein amplified by RT-PCR from mouse testis RNA was cloned into the pEGFP-C1 vector to form the recombinant PRP-pEGFP-C1 plasmid.Then the recombinant plasmid was transfected into 293FT cells

and the expression of PRP was detected by RT-PCR and Western blotting.MTT and flow cytometry(FCM)analysis were employed to analysis the cell proliferation activity and the cell cycle

respectively.Results:PRP coding region was successfully cloned into the eukaryotic vector pEGFP-C1.The recombinant vector was transfected into 293FT

results of RT-PCR and Western blotting showed that the PRP could express in 293FT cells.Overexpressing PRP gene decreased the cell proliferation activity as well as disturbing the cell cycle distribution of 293FT cells in vitro.Conclusion:We have constructed a recombinant vector expressing PRP and eGFP fusion protein

which is in favour of the further study on the gene function of PRP.In addition

PRP has anti-proliferin activity in human-derived cells through disturbing the cell cycle

suggesting that PRP maybe act as a potential gene therapeutic candidate used in human tumor

not just mouse tumor.

关键词

proliferin related proteinpEGFP-C1细胞增殖

Keywords

proliferin related proteinpEGFP-C1cell proliferation

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