%0 Journal Article
%T Expression and Purification of ATP Sulfurylase from Saccharomyces cerevisias in Escherichia coli and Its Application in Pyrosequencing<br>酿酒酵母ATP-硫酸化酶在大肠杆菌中的表达纯化及其在焦测序中的应用
%A LUO Juan
%A WU Wen-Juan
%A ZOU Bing-Jie
%A ZHOU Guo-Hua
%A <br>罗娟
%A 吴文娟
%A 邹秉杰
%A 周国华
%J 生物工程学报
%D 2007
%I 
%X ATP sulfurylase (ATPS,EC 2.7.7.4) reversibly catalyzes the reaction between ATP and sulfate to produce APS and pyrophosphate (PPi), and has been used in pyrosequencing. The gene coding ATP sulfurylase was amplified from the genomic DNA of Saccharomyces cerevisias (CICC 1202), and cloned into prokaryotic expression plasmid pET28a( + ) to provide a recombinant expression plasmid pET28a( + )-ATPS. Upon IPTG induction, ATP sulfurylase was produced by E. coli BL21 (DE3) harboring the recombinant expression plasmid pET28a( + )-ATPS. The relative molecular weight of recombinant ATP sulfurylase with His tag was about 60 kD. The recombinant ATP sulfurylase with electrophoretic pure grade was obtained only by two purification steps: His * Bind Resin affinity chromatography and ultrafiltration. The specific activity of the purified recombinant ATP sulfurylase was as high as 5.1 x 10(4) u/mg. The successful application of the enzyme in pyrosequencing was also demostrated.
%K ATP sulfurylase
%K Saccharomyces cerevisias
%K expression
%K purification
%K pyrosequencing<br>ATP-硫酸化酶
%K 酿酒酵母
%K 表达
%K 纯化
%K 焦测序
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=D98BF98D3F90A5C9&yid=A732AF04DDA03BB3&vid=EA389574707BDED3&iid=E158A972A605785F&sid=F8AEC975DBDD7F2F&eid=821800203AD09E7B&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=0&reference_num=15