%0 Journal Article
%T Cloning and Expression of Extracellular Domain of Prostate Specific Membrane Antigen in Escherichia coli and Preparation of Polyclonal Antibody<br>人前列腺特异膜抗原膜外区基因的克隆表达及抗体的制备
%A YE Chuan-Zhong
%A ZHAO Xu-Dong
%A ZHANG Fang-Lin
%A LIN Zhen XU Ming
%A ZHANG Yong-Kang
%A CHEN Chang-Qing
%A <br>叶传忠
%A 赵旭东
%A 张芳林
%A 林震
%A 许明
%A 张永康
%A 陈常庆
%J 生物工程学报
%D 2002
%I 
%X Human Prostate Specific Membrane Antigen(PSMA) cDNA was amplified using total RNA extracted from prostate carcinoma tissue by RT-PCR. The cDNA fragment of extracellular domain of PSMA(edPSMA) gene was amplified by PCR and cloned into expression vector pMAL-c2x. Sequence analysis of both PSMA and edPSMA revealed identity to the GenBank reported. The edPSMA was expressed in E. coli as part of a fusion protein with MBP as the induction of IPTG. Western blot analysis showed the recombinant protein could react with PSMA monocloned antibodies 4G5. MBP-edPSMA fusion protein were purified by amylose resin affinity chromatography and showed to be homogeneity in SDS-PAGE(120 kD). BALB/C mice were immunized with the purified protein for the preparation of polyclonal antibody. The polyclonal antibody, which had a title of 1:12,800, were indicated the specificity to prostate tissue.
%K extracellular domain of prostate-specific membrane antigen
%K expression
%K affinity chromatography
%K fusion protein
%K polyclonal antibody<br>前列腺特异膜抗原膜外区
%K 原核表达
%K 亲和层析
%K 抗体
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=B2E29676334CE134&yid=C3ACC247184A22C1&vid=13553B2D12F347E8&iid=CA4FD0336C81A37A&sid=6209D9E8050195F5&eid=7C3A4C1EE6A45749&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=0&reference_num=8