%0 Journal Article
%T Expression of PPDK from Microbispora rosea subsp. aerata in Escherichia. coli and Its Application in Pyrosequencing<br>热玫瑰小双孢菌来源的丙酮酸磷酸双激酶的表达及应用
%A Bingjie Zou
%A Zhiyao Chen
%A Guohua Zhou
%A <br>邹秉杰
%A 陈之遥
%A 周国华
%J 生物工程学报
%D 2008
%I 
%X Pyruvate phosphate dikinase (PPDK; EC 2.7.9.1) is found in certain microorganisms and plants, and catalyzes the conversion of AMP, PPi and phosphoenolpyruvate (PEP) to ATP, Pi and pyruvate. Using the genomic DNA of Microbispora rosea subsp. aerata as the template, a DNA fragment encoding the gene PPDK was amplified by PCR and inserted into the expression vector pET28a(+), yielding pET28a (+)-PPDK. The E. coli BL21 (DE3) was transformed with the pET28a (+)-PPDK. After inducing with IPTG, the E. coli BL21 (DE3) pET28a (+)-PPDK] expressed recombinant PPDK fused to an N-terminal sequence of 6-His Tag. The molecular weight of PPDK was estimated to be 101 kD by SDS-PAGE. The PPDK was purified by His ? Bind Resin affinity chromatography and ultrafiltration using 10 kD cut-off membrane. The successful application of PPDK in pyrosequencing was also demonstrated.
%K PPDK
%K Microbispora rosea subsp
%K aerata
%K E
%K coli
%K pyrosequencing<br>PPDK
%K 热玫瑰小双孢菌
%K 大肠杆菌
%K 焦测序
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=FBD1B15B9F5CA7682A1F6B239A1E661F&yid=67289AFF6305E306&vid=B91E8C6D6FE990DB&iid=E158A972A605785F&sid=DC06EBDBAF4E06D3&eid=21A4BC96BDC43D33&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=0&reference_num=15