| 摘要: |
| 通过原核表达系统表达猪链球菌Sao-M和IdeSsuis基因,并分析其反应原性。根据GenBank数据库发表的序列,利用分子生物学软件设计了2对特异性引物,通过PCR扩增Sao-M和IdeSsuis基因,经测序分析后,分别克隆到pET28a(+)和pMAL-c2X,构建重组表达质粒pET28a:Sao-M和pMAL-c2X:IdeSsuis,然后将鉴定为阳性的重组质粒分别转入宿主菌BL21和Rosetta2中用IPTG进行诱导表达,同通过免疫印迹进行鉴定。实验表明,两种重组蛋白在原核系统中获得了高效的表达,并能与相应的阳性抗血清发生特异性反应,本研究为进一步探究Sao-M和IdeSsuis蛋白生物学功能及其在猪链球菌致病机制上的作用提供了依据。 |
| 关键词: 猪链球菌 Sao-M蛋白 IdeSsuis蛋白 原核表达 反应原性 |
| DOI： |
| 投稿时间：2016-04-02修订日期：2016-06-02 |
| 基金项目: |
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| Expression and Western blotting Analysis of Streptococcus suis Sao-M and IdeSsuis |
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| (Shandong Binzhou Animal Science Veterinary Medicine Academy,Binzhou) |
| Abstract: |
| Prokaryotic expression system was used to express Sao-M and IdeSsuis of Streptococcus suis (S.suis), and the immunogenicity was identified. In order to obtain the Sao-M and IdeSsuis complete open reading frame sequence, based on comparison of sequences reported in the GenBank database, two pairs of specific primers were designed to amplify those two genes by using polymerase chain reaction (PCR) and the product of PCR was sequenced and subcloned into pET28a( ) and pMAL-c2X respectively, getting two recombinant plasmids named pET28a:Sao-M and pMAL-c2X:IdeSsuis. Recombinant plasmids were transformed into BL21 and Rosetta 2 and induced by IPTG. Western-blotting was used for testing the recombinant proteins. The results showed that recombinant proteins were highly expressed, and could react with ployclonal antibody. |
| Key words: Streptococcus suis Sao-M IdeSsuis prokaryotic expression reactogenicity |
