The construction and significance of maspin/PCR2.1 expression vector

YAN Xiong; et al

doi:10.13406/j.cnki.cyxb.2005.03.020

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The construction and significance of maspin/PCR2.1 expression vector

更新时间：2024-01-17

- The construction and significance of maspin/PCR2.1 expression vector
- Journal of Chongqing Medical University Issue 3, Pages: 379-382(2005)
- 作者机构：
  
  1. 重庆医科大学临床学院普外科
  2. 重庆医科大学临床学院普外科,重庆,400016
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2005.03.020
  CLC： R735.7
- Published：2005，
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[1]阎雄,姚榛祥.maspin/PCR2.1表达载体的构建及意义[J].重庆医科大学学报,2005(03):379-382.

YAN Xiong, et al. The construction and significance of maspin/PCR2.1 expression vector. [J]. Journal of Chongqing Medical University (3):379-382(2005)
[1]阎雄,姚榛祥.maspin/PCR2.1表达载体的构建及意义[J].重庆医科大学学报,2005(03):379-382. DOI： 10.13406/j.cnki.cyxb.2005.03.020.

YAN Xiong, et al. The construction and significance of maspin/PCR2.1 expression vector. [J]. Journal of Chongqing Medical University (3):379-382(2005) DOI： 10.13406/j.cnki.cyxb.2005.03.020.

摘要

目的:构建maspin/PCR2.1表达载体

为深入研究maspin基因抑制肝癌生长、转移、浸润作用和机制奠定基础。方法:PCR扩增maspin基因全长

将表达载体PCR2.1用Hind111+XbaI进行双酶切后

T4DNA连接酶连接成重组质粒

转染到人肝癌高转移细胞株MHCC-97中进行表达

检测转染前后maspin表达的变化。结果:重组质粒在大肠杆菌株JM109内扩增;提纯、纯化后

用HindIII、XbaI酶切鉴定及测序鉴定证明maspin基因已完整、正确的插入到PCR2.1表达载体

并在人肝癌MHCC-97细胞中上调了maspin基因mRNA和蛋白水平表达。结论:成功构建了maspin/PCR2.1表达载体

能在真核细胞中。

Abstract

Objective:To construct the expression vector maspin /Pcr2.1 in order to provide the evidence for the inhibitory effect of maspin gene on growth

metastasis and invasion of hepatocellular carcinoma(HCC) and to discuss its mechnism.Methods:The full length of maspin gene was cloned

and the expression vector PCR2.1 was digested by Hind111/XbaI

then the maspin gene and the PCR2.1 vector were ligated with T4 DNA ligase to be recombinant vector.The recombinant vector maspin/PCR2.1 was transfected into humman hcc MHCC-97 cell line

and the expressive changes of maspin gene were detected.Results:The recombinant plasmid was amplified in the E.coli.JM109.After the identification of redigesting by HindIII/XbaI and sequencing

the reconstructive plasmid was confirmed to contain the correct and full nucleotide sequence of maspin gene

and the mRNA and protein level of maspin gene were up-regulated in hcc MHCC-97 cell.Conclusion:The expression vector maspin/PCR2.1 is constructed successfully and can be expressed in eukaryotic cells.

关键词

maspinPCR2.1表达载体构建意义

Keywords

maspinPCR2.1 expression vectorconstructionSignificance

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