%0 Journal Article
%T Expression and Characterization of Envelope Protein 2 Gene of Hepatitis G Virus in Pichia pastoris<br>庚型肝炎病毒E2区cDNA在毕赤酵母中的表达及抗原性鉴定
%A WANG Zhuo
%A |Hua
%A YE Kai
%A XU Hong
%A MA Hui
%A |Wen
%A <br>王卓华
%A 叶凯
%A 徐洪
%A 马辉文
%A 童立恒
%A 彭习亮
%J 生物工程学报
%D 2002
%I 
%X A cDNA fragment locating at the putative envelop protein 2(E2) region of GBV-C/HGV fused with Schistosoma japonicum, glutathione S-transferase(GST) was amplified with PCR from plasmid pGEX-E2. The amplified DNA fragment was inserted into plasmid pGEX-5X-1, at the downstream of the coding sequences of GST, in the same reading frame with the gene of GST. The fusion gene fragment of GST-E2 was amplified with PCR, using the recombinant plasmid pGEX-5X-1-E2 as the template. The amplified 1324 bp DNA fragment of GST-E2 was inserted into Pichia pastoris expression vector pPIC9K in reading frame with alpha-factor secreting signal peptide. The plasmid pPIC9K-GST-E2 was transformed into Pichia pastoris GS115 with electroporation. The transformants (His+ Muts) were selected and induced to express the 54kD GST-E2 fusion protein, which could be specially recognized by both the antisera directed against E2 and against GST. The GST-E2 fusion protein was purified with Sepharose 4B glutathione affinity chromatography to a purity of 95%. The expression was optimized to achieve the highest expression level of GST-E2 fusion protein which was accumulated up to 50% of total proteins in the culture supernatant. The GST-E2 protein derived from the recombinant Pichia pastoris was proved possessing antigenicity and high specificity by ELISA, probed with sera from the patients infected by GBV-C/HGV.
%K GBV\|C/HGV
%K E2 protein
%K Pichia pastoris
%K expression
%K antigenicity<br>庚型肝炎病毒，
%K E2膜蛋白，
%K 巴斯德毕赤氏酵母，
%K 表达，
%K 抗原性
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=1BDC33EE59026E88&yid=C3ACC247184A22C1&vid=13553B2D12F347E8&iid=0B39A22176CE99FB&sid=3E0812ED84A7B31D&eid=C29816B2656377A7&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=2&reference_num=12