%0 Journal Article
%T Construction of Tandem Fragment Polymer to Generate Polyclonal Antibody for Recognizing Plasmid PRSET-A expressed Peptides<br>质粒pRSET-A前导肽串联多聚体的构建及其多克隆抗体制备*
%A HUANG Yun-mao
%A YUE Guang-fang
%A LIU Li
%A LIU Ying
%A LI Wan-li
%A SHI Zhen-dan
%A <br>黄运茂
%A 岳光芳
%A 刘丽
%A 刘颖
%A 李万利
%A 施振旦
%J 中国生物工程杂志
%D 2008
%I 
%X The plasmid pRSET-A, a common prokaryotic expression vector, encodes a N-terminal 34aa precursor peptide with a histidine tag sequence. Anti-histidine-tag antibody is usually used for identification of expression of the recombinant protein expressed using pRSET-A. A pair of primers that contained flanking hydropathic amino acid codons were synthesized, to amply the sequence coding for 10~34 aa in the precursor sequence. The amplified fragment was inserted into pRSET-A to generate the first double repeat of the precursor gene. By utilizing a pair of isocaudamer BamH I and Bgl II sites, and another downstream Hind III site of plasmid pRSET-A, following a series of simple double digestions and ligation of the resulted products, a series of repeat (3, 4 and 6) precursor peptide fragment genes were derived. The 6 repeat polymer of the precursor peptide was successfully expressed, and used to immunize goats for preparation of anti-precursor peptide antibodies. The antibody produced effectively identified the recombinant protein with the completed precursor peptide, but not the recombinant protein only had the His-tag sequence and lack the 10~34 aa sequence. These results demonstrated the repeated short-peptides polymer can be quickly and efficiently constructed by utilizing a pair isocaudamer sites in production of subunit-vaccine or immune-regulating antigens.
%K Plasmid
%K Repeat fragment polymer
%K Antibody
%K Preparation
%K Application<br>质粒
%K 串联多聚体
%K 抗体
%K 制备
%K 应用
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=951380788B355DEA7D75520FA3B199C9&aid=E7BB6F031901461C7E5BB88EFF0F5319&yid=67289AFF6305E306&vid=D3E34374A0D77D7F&iid=9CF7A0430CBB2DFD&sid=E44E40A2398D4F2A&eid=0D0D661F0B316AD5&journal_id=1671-8135&journal_name=中国生物工程杂志&referenced_num=1&reference_num=12