Building and identification of prion gene eukaryotic expression vector of human
10.3969/j.issn.1671-8348.2014.19.013
- VernacularTitle:人朊蛋白基因真核表达载体的构建及鉴定
- Author:
Haiying ZHANG
;
Yiheng LIU
;
Xiaowen FEI
;
Xinan YI
;
Duoqing WU
- Publication Type:Journal Article
- Keywords:
alzheimer′s disease;
PRNP gene;
pEGFP-N2 eukaryotic expression vector
- From:
Chongqing Medicine
2014;(19):2434-2436
- CountryChina
- Language:Chinese
-
Abstract:
Objective To constructan eukaryotic expression recombinant plasmid named pEGFP-N2-PRNP .Methods Total RNA was extracted from alzheimer (AD) disease peripheral blood ,and the PRNP gene was amplified by reverse transcription-poly-merase chain reaction(RT-PCR) .By using gene recombination technique ,human PRNP cDNA was inserted into retroviral vector pEGFP-N2 .The recombinant plasmid was identified by a pair of specified primers containing the restriction sites of Xho Ⅰ and EcoRⅠ .Results The PRNP gene could be obtained by RT-PCR ,the recombinant plasmid was identified by restriction endonucle-ase analysis ,PCR and sequence analysis ,and the expression vector pEGFP-N2-PRNP ,which could be stably expressed in SH-SY5Y cells .Conclusion The recombinant plasmid pEGFP-N2-PRNP is constructed successfully ,Which offers a basic for the further re-search on PRNP biological fuction .
