逆轉(zhuǎn)錄病毒載體plxpxsntcrα122iresvβ7.1的構(gòu)建及其殺傷肝癌細(xì)胞作用的研究

1、廣東藥學(xué)院碩士學(xué)位論文逆轉(zhuǎn)錄病毒載體pLXPXSN-TCRα12-2-IRES-Vβ7.1的構(gòu)建及其殺傷肝癌細(xì)胞作用的研究姓名：牟艷芳申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：病原生物學(xué)指導(dǎo)教師：黃樹林20090501廣東藥學(xué)院碩士研究生論文 iConstruction of Recombinant Retrovirus Vector pLXPXSN- TCRα12-2-IRES-Vβ7.1 and Study of the Cytotoxicity t

2、o Hepatoma Cell Graduate: MOU Yan-fang Major: Biological Etiology Supervisor: Professor Huang Shu-lin Abstract Objective To construct a dicistronic retrovirus expression vector to coxpress both human TCRα12-2 and TCRVβ

3、7.1genes and study the cytotoxicity to hepatoma cell. The use of Recombinant Retrovirus Vector pLXPXSN-TCRα12-2-IRES-Vβ7.1 could perhaps be potent tools for cancer gene therapy. Methods 1, construct the plasmid pLXPXSN-

4、TCRα12-2-IRES-Vβ7.1. The TCRα12-2 and TCRVβ7.1 were amplified by PCR from plamid that we constructed before，which was cloned into vector pLXPXSN .2, package and identification of the recombinant retrovirus pLXPXSN-TCRα1

5、2-2-IRES-Vβ7.1. The plasmid pLXPXSN-TCRα12-2-IRES-Vβ7.1 was transfered into PA317 cells using Lipofectamine 2000 to generate recombinant retrovirus.A replication-defective retrovirus pLXPXSN-TCRα12-2-IRES-Vβ7.1 cou

6、ld be made by homologous recombinant,which can simultaneously express TCRα12-2 gene and TCRVβ7.1 gene.The recombinant retrovirus was analyzed by PCR to test target genes;The expression of pLXPXSN- TCRα12-2-IRES-Vβ7.1 i

7、n Hela cells was confirmed by reverse transcription polymerase chain reaction procedure(RT-PCR).The protein TCRVβ7.1 was detected by flow cytometry in PBMCs which were infected by pLXPXSN-TCRα12-2-IRES-Vβ7.1.The recombi