%0 Journal Article
%T Construction of MAGE-3 Prokaryotic Expression Plasmid and Its Expression in Escherichia coli<br>MAGE-3原核表达载体的构建和表达
%A SUN Xiao Dong
%A WU Jin Min
%A LIU Xing E
%A <br>孙晓东
%A 吴金民
%A 刘杏娥
%J 生物工程学报
%D 2003
%I 
%X To express the GST MAGE 3 protein in E.coli , and investigate the antitumor immune responses induced by Dendri tic cells(DCs) pulsed with GST MAGE 3 protein, the recombinant expression plasmid pGEX MAGE 3 was constructed by ligating MAGE 3 gene, which was amplified by RT PCR and confirmed by sequencing, and the pGEX 4T 2 vector. The recombinant plasmid was transformed into BL 21 E.coli . The expression of GST MAGE 3 was induced with IPTG. The GST MAGE 3 protein expressed as high as 32% of the total cellular protein. After purification with Glutathione Sepharose 4B, the purity of the protein was more than 90%, and 3mg GST MAGE 3 was obtained from 100 mL BL 21 lysate. Dendritic cells from gastric carcinoma patients were pulsed with GST MAGE 3 protein, and these DCs were used to stimulate the autologous T lymphocytes. After 7 days, the T lymphocytes cocultured with DCs pulsed with GST MAGE 3 antigen exhibited specific cytotoxicity against MAGE 3 positive SGC 7901 cells. It is concluded that the GST MAGE 3 protein are able to present antigen to T lymphocytes, activate antigen specific CTLs and induce special antitumor immune responses in vitro . Our results lay the groundwork for further research of the MAGE 3 vaccine.
%K GST
%K MAGE
%K 3 protein
%K prokaryotic expression
%K dendritic cells(DCs)
%K cytotoxic T lymphocytes (CTLs)
%K immune
%K response<br>GSTMAGE-3蛋白，
%K 原核表达，
%K 树突状细胞，
%K 细胞毒T淋巴细胞，
%K 免疫应答
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=0D380837DDE21687&yid=D43C4A19B2EE3C0A&vid=2A8D03AD8076A2E3&iid=38B194292C032A66&sid=CAA7BAE04CB631A1&eid=5D8C08279A19B0D4&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=1&reference_num=14