| 摘要: |
| 根据家蝇幼虫防御素基因（Mdd I）序列，设计合成含有Kpn I和Xba I酶切位点的特异性引物，PCR扩增出Mdd I基因全长序列。克隆并测定序列后，构建真核表达质粒pGAPZαA-Mdd I，将其在毕赤酵母（P. pastoris）GS115中分泌表达，并对表达产物进行抑菌活性分析。结果表明，Mdd I基因在P. pastoris中成功表达，分子量约为10.3kDa，抑菌结果显示Mdd I基因的真核表达产物对猪源链球菌有抑制作用。该研究成功构建了表达Mdd I基因的P. pastoris菌株，Mdd I基因在真核表达系统中的成功表达为进一步研究表达产物的生物学活性和免疫学活性奠定了基础。 |
| 关键词: 家蝇 防御素 P. pastoris 真核表达 抗菌活性 |
| DOI： |
| 投稿时间：2015-07-05修订日期：2015-08-01 |
| 基金项目:国家自然科学基金资助项目(31140026)；吉林省世行贷款农产品质量安全项目(2011-Y05) |
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| Expression of Musca domestica Larvae Defensin Gene MddⅠin P.pastoris and Identification of its Antibacterial Activity |
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| (Jilin Agricultural University) |
| Abstract: |
| A pair of primers with restriction sites Kpn I and Xba I were designed and synthesized according to the gene sequence of Musca domestica larvae defensin gene Mdd I, then facilitated to amplify the full-length sequence of Mdd I by polymerase chain reaction (PCR). After cloning and sequencing, the eukaryotic expression plasmid pGAPZαA-Mdd I was constructed and then expressed in P. pastoris GS115, subsequently the antimicrobial activity of recombinant protein was analyzed. The result showed that the expressed protein of Mdd I was about 10.3KDa, which possessed antibacterial activity against swine Streptococcus. Conclusively, the recombinant P.pastoris with gene Mdd I was successfully constructed, which laid the foundation for research and development of Musca domestica defensin gene as antibacterial feed additives. |
| Key words: Musca domestica defensin P. pastoris eukaryotic expression antimicrobial activity |
