%0 Journal Article
%T Expression And Purification Of A Novel Influenza Virus Subunit<br>一种新型广谱流感病毒亚单位疫苗的表达与纯化
%A WEI Yi-ju
%A LONG Hai-ting
%A YANG Xu
%A LI Jian-fang
%A BI Yan-wei
%A LI Jian-feng
%A XU Wei-ming
%A <br>魏义举
%A 龙海亭
%A 杨旭
%A 李建芳
%A 毕研伟
%A 李健峰
%A 徐维明
%J 中国生物工程杂志
%D 2007
%I 
%X The influenza A virus matrix protein2 gene(M2) which deleted transmembrane region was amplified by overlap extending PCR, and the multi-epitope gene of hemagglutinin(HA) was PCR amplified with seven continuous synthesized segments by designing primer. The two gene segments were separately cloned into pMD18-T vector to sequence analysis and prokarytic expression vector pET28a+ to construct the recombinant plasmid pET28a+-M2d-HA. The recombinant plasmid was transformed into E.coli BL21(DE3), and the high expression strain was obtained by screening monoclones. The recombinant protein existed as inclusion bodies, which accounted about 45% of the total cellular protein. The inclusion bodies were washed with 1% Triton X-100 solution twice, and dissolved in 8 mol/L urea solution. The solution protein was purified by Ni2+ affinity chromatography, and refolded by dilution renaturation, then purified by Q Sepharose FF cation exchange column. The purity of the protein was over 90% by HPLC analysis. The result of western-blot showed it has good antigenicity and specificity. These results strongly supported for the further study of the broad-spectrum influenza virus subunit vaccine.
%K 流感病毒
%K 亚单位疫苗
%K 多表位
%K 表达纯化
%K 抗原性
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=951380788B355DEA7D75520FA3B199C9&aid=E5A3A9FD00FD8F09&yid=A732AF04DDA03BB3&vid=DB817633AA4F79B9&iid=94C357A881DFC066&sid=869807E2D7BED9EC&eid=C36EC077A8A90308&journal_id=1671-8135&journal_name=中国生物工程杂志&referenced_num=0&reference_num=13