%0 Journal Article
%T Cloning and Expression of Pokeweed Antiviral Protein-Ⅱ Gene from the Summer Leaves of Phytolacca amercana<br>美洲商陆抗病毒蛋白-Ⅱ基因的克隆和表达
%A HUANG Jian-Song
%A ZHAN Jin-Biao
%A ZOU Yuan
%A FENG Wei-Hong
%A <br>黄建松
%A 詹金彪
%A 邹媛
%A 冯微宏
%J 生物工程学报
%D 2006
%I 
%X The cDNA sequence encoding pokeweed antiviral protein-II was cloned from the fresh summer leaves of phytolacca amercana by RT-PCR. The recombinant PAP-II was subcloned into the expression vector pET-28a(+) and expressed in E. coli BL21 after IPTG induction. SDS-PAGE analysis showed that the expressed PAP-II existed in the form of inclusion bodies. The purified fusion protein was obtained after a series of steps including cell break, inclusion body solubilization, protein refolding and purification through BBST NTA resin column. The non-radioactive ELISA-based HIV-1 integrase assay showed that the recombinant pokeweed antiviral protein-II and RTA were able to inhibit HIV-1 integrase to some extent (IC50 = 303 microg/mL, 220 microg/mL respectively). MTT assay showed that cytotoxicity of pokeweed antiviral protein II for HEP-G2 cells and Hela cells was in a dose-dependent manner with IC50 s of 93 microg/mL and 102 microg/mL, respectively. The results suggested that pokeweed antiviral protein-II is a potent anti-tumor candidate. The finding of integrase inhibitory activity and the discovery of cytotoxicity provide more insights into the anti-HIV and the anti-tumor activities of PAP-II.
%K 核糖体失活蛋白
%K 美洲商陆抗病毒蛋白-Ⅱ
%K HIV-1整合酶
%K 表达
%K 细胞毒性
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=EEC53A17D505252C&yid=37904DC365DD7266&vid=BC12EA701C895178&iid=E158A972A605785F&sid=106103EB0EA31435&eid=0A8675156EB60B87&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=3&reference_num=17