%0 Journal Article
%T Construction of a multi-copy Pichia expression vector and expression of human brain-derived neurotrophic factor in Pichia pastoris<br>一种多拷贝毕赤酵母表达载体的构建及人脑源性神经营养因子的表达
%A LIANG Wei-feng
%A ZHANG
%A Chao-chun
%A YANG Xi-cai
%A <br>梁伟锋
%A 张朝春
%A 杨希才
%J 微生物学报
%D 2005
%I 
%X A pPICZ alpha1 vector was reconstructed with pPICZ alpha vector and PCR product of His4-Kan sequence from pPIC9K vector. The sequence encoding human brain-derived neurotrophic factor (hBDNF) was inserted into pPICZ alpha1. The expression vectors carrying 1, 2, 3, 6 copies of expression cassettes were constructed and transformed into Pichia pastoris GS115 respectively. The transformants were screened with G418/Zeocin and induced by 0.5% methanol, and the results of SDS-PAGE confirmed that the expression protein was about 14 kD and the percentage of the protein was about 20% of the total secreted protein. ELISA and Western blot analysis demonstrated that the protein could be specifically combined with chicken antibodies to human-BDNF.
%K Multi-copy Pichia expression vector
%K hBDNF
%K Pichia pastoris
%K Expression<br>多拷贝表达载体
%K 人脑源性神经营养因子
%K 毕赤酵母
%K 分泌表达
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=13B1729ECC9B6B9B&yid=2DD7160C83D0ACED&vid=94E7F66E6C42FA23&iid=CA4FD0336C81A37A&sid=339D79302DF62549&eid=16D8618C6164A3ED&journal_id=0001-6209&journal_name=微生物学报&referenced_num=5&reference_num=11