%0 Journal Article
%T Cloning of PTD-NPY Fusion Gene and Its Secretory Expression in Pichia pastoris<br>PTD-NPY融合基因的克隆及其在毕赤酵母中的分泌表达
%A Yucheng Sun
%A Fengqiu Zhou
%A Jiayu Wan
%A Hongwei Gao
%A <br>孙玉成
%A 周凤秋
%A 万家余
%A 高宏伟
%J 生物工程学报
%D 2008
%I 
%X The PTD-NPY fusion gene derived from HIV-1 TAT protein transduction domain and rat neuropeptide Y was amplified by overlap extension PCR, digested and subcloned into yeast expression vector pPICZaA to construct recombinant expression plasmid pPICZa-PTD-NPY. The cloned PTD-NPY fusion gene was identified by PCR and restriction enzyme digestion and sequenced. The exact recombinant plasmid was linearized by Sac I and integrated by electrotransformation into the genome of Pichia pastoris GS115 cells. Then, these positive recombinant yeast cells were induced by 10 mL/L methanol to express soluble PTD-NPY fusion protein. After 120 h of methanol induction, the SDS-PAGE electrophoresis result indicated PTD-NPY fusion protein was efficiently secreted into the medium. Western blotting analysis proved that the expressed fusion protein had specific NPY binding activity. The successful expression of PTD-NPY fusion protein in Pichia pastoris provided basis for its further application study.
%K protein transduction
%K neuropeptide Y (NPY)
%K fusion gene
%K cloning
%K secretory expression<br>蛋白转导
%K 神经肽Y(NPY)
%K 融合基因
%K 克隆
%K 分泌表达
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=EBF43FBE14BA340B1C3B9789EC2719B9&yid=67289AFF6305E306&vid=B91E8C6D6FE990DB&iid=9CF7A0430CBB2DFD&sid=28132669ED57E050&eid=967EC9935A34C4F3&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=1&reference_num=9