%0 Journal Article
%T A Pichia pastoris with α-1, 6-mannosyltransferases Deletion and Its Use in Expression of HSA/GM-CSF Chimera<br>α-1,6-甘露糖转移酶基因敲除的毕赤酵母菌株构建及其用于融合蛋白HSA/GM-CSF 表达的研究
%A WANG Yue
%A GONG Xin
%A CHANG Shao-Hong
%A LIU Bo
%A SONG Miao
%A HUANG Hai-Hua
%A WU Jun
%A <br>王越
%A 巩新
%A 唱韶红
%A 刘波
%A 宋淼
%A 黄海华
%A 吴军
%J 生物工程学报
%D 2007
%I 
%X Yeast is a widely used host for recombinant protein expression. However, glycoproteins derived from yeast contain N-glycan of high mannose type and are usually hyperglycosylated. alpha-1,6-mannosyltransferases gene (och1) encodes the enzyme that initiates the first step of out-chain elongation of high mannose type N-glycan in yeast, which is different from that in human. So, a high efficient method to knockout target gene by two-step recombination was established and was used to delete och1. In the first recombinant, a plasmid with och1::ADE1 and ura3 gene was linearized in the downstream of och1 and inserted to the och1 site of P. pastoris genome, where the upstream and downstream of och1 were duplicated. In the second recombinant, the duplicated fragments of och1 were exchanged and the och1 deletion strains were selected on the plates containing 5-FOA, but no adenine. Then the och1 deletion strain was applied to express an human serum albumin (HSA) granulocyte-macrophage colony-stimulating factor (GM-CSF) chimera. Different with the hyperglycosylated HSA/GM-CSF chimera expressed in wild type P. pastoris, the chimera expressed in the och1 deletion strain, contained smaller N-glycan. The results suggested that the och1 mutant yeast may be more suitable for production of recombinant glycoproteins. And the och 1 deletion strain could be used for further re-engineering to produce complex human glycoproteins.
%K α-1
%K 6-甘露糖转移酶
%K 基因敲除N-糖基化
%K 粒细胞-巨噬细胞集落刺激因子
%K 人血清白蛋白
%K 毕赤酵母
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=2C45CBBF599FA9A0&yid=A732AF04DDA03BB3&vid=EA389574707BDED3&iid=94C357A881DFC066&sid=3382A18868551611&eid=FE68044BBD4BDA5F&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=0&reference_num=18