%0 Journal Article
%T Cloning Expression and Purification of Glycerol Dehydrogenase from Klebsiella pneumoniae<br>克雷伯杆菌甘油脱氢酶基因的克隆表达与纯化
%A Tingting Zhang
%A Baishan Fang
%A Geng Wang
%A Feifei Wang
%A <br>张婷婷
%A 方柏山
%A 王耿
%A 王飞飞
%J 生物工程学报
%D 2008
%I 
%X The gldA gene coding glycerol dehydrogenase (GDH) was amplified by PCR with the genomic DNA of Klebsiella pneumoniae as the template. The gldA were inserted in pMD-18T to construct the recombinant cloning vector pMD-gldA. After the DNA sequence was determined, the gldA was subcloned into expression vector pET-32a (+) to construct the recombinant expression vector pET-32gldA. Upon lactose induction, soluble GDH was over-produced by E. coli BL21 (DE3) harboring the expression construct. Recombinant GDH purified by Ni-NTA affinity chromatography showed a single band about 54 kD on SDS-PAGE gel, and the specified activity was about 188 u/mg, the purification fold is 3 times and the activity recovery is 67.5%.
%K glycerol dehydrogenase
%K Klebsiella pneumoniae
%K protein purification<br>甘油脱氢酶(GDH)
%K 克雷伯杆菌
%K 蛋白纯化
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=08E7B2E74D0FAEA49176DF2D04611781&yid=67289AFF6305E306&vid=B91E8C6D6FE990DB&iid=38B194292C032A66&sid=DDDA4F26E8AD3C0E&eid=A03A15CF5604A8B0&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=1&reference_num=13