%0 Journal Article
%T Prokaryotic expression of OC-IdeltaD86 (Oryzacystatin-IdeltaD86) gene and analysis of its activity<br>OC-IΔD86 (Oryzacystatin-IΔD86)基因的原核表达及活性分析
%A Yumeng Huo
%A Qiwei He
%A Shuangyi Zhao
%A Yuanfang Xu
%A <br>霍雨猛
%A 何启伟
%A 赵双宜
%A 徐苑芳
%J 生物工程学报
%D 2008
%I 
%X According to the amino acids sequence of OC-IdeltaD86 gene and Escherichia coli codon usage, we synthesized this gene by overlap extension PCR method with 7 oligonucleotides DNA fragments. The PCR fragment was inserted into pGEM-T-easy vector and the recombined plasmid was named pGEM-T-OC-IdeltaD86. Two oligonucleotides into which the BamH I and Xho I sites were introduced were designed and synthesized based on pGEM-T-OC-IdeltaD86 and pet21b, and the PCR fragment into which the BamH I and Xho I sites were introduced was obtained. After digesting it with BamH I and Xho I, OC-IdeltaD86 gene was cloned into the corresponding sites of pet21b and obtained prokaryotic expression vector pet21b-OC-IdeltaD86. OC-IdeltaD86 gene was expressed in E. coli (BL21(DE3)plysS) after IPTG(Isopropyl beta-D-1-thiogalactopyranoside) inducement for 5 hours. The fusion protein of OC-IdeltaD86:6His gene accounted for 11.4% of total protein and 16.4% of soluble protein, which had been successfully purified by Ni-NTA and concentrated by PEG20000. This protein can effectively inhibit papain activity in vitro and may be used in anti-nematode research in vivo.
%K OC-IΔD86
%K 重叠延伸PCR
%K 抑制剂
%K 纯化
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=EAFB3D9A3E23F572590AAB4FC1A6AB0D&yid=67289AFF6305E306&vid=B91E8C6D6FE990DB&iid=DF92D298D3FF1E6E&sid=1805714CB338690A&eid=9978E8308B7F7CE6&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=0&reference_num=0