| 摘要: |
| 克隆奶牛乳房炎金黄色 葡萄球菌裂解性噬菌体裂解酶LysIMEP5基因,分析其生物信息学特性。以本实验室分离的奶牛乳房炎金黄色葡萄球菌裂解性噬菌体vB_SauS_IMEP5 为材料,根据其全基因组学信息,获取裂解酶基因序列,应用Primer 5.0设计特异性引物,采用PCR方法扩增并克隆LysIMEP5基因,通过BLAST进行序列比对分析,利用在线软件对蛋白结构进行预测。成功克隆到裂解酶LysIMEP5基因,测序结果通过DNAMAN比对与原序列完全匹配,无任何基因突变,同源性分析显示,其已报道的金黄色葡萄球菌噬菌体裂解酶相似性最高为83.1% ；裂解酶LysIMEP5 基因序列全长1371bp,共编码456个氨基酸,为亲水性蛋白,分子质量为51.717 kDa,理论等电点为9.70； LysIMEP5同时存在CHAP片段和Amidase-3片段；该蛋白无跨膜区,无信号肽,以无规则卷曲为主。从奶牛乳房炎金黄色葡萄球菌裂解性噬菌体中成功克隆出LysIMEP5基因,通过对该蛋白的结构预测分析为后续克隆表达,及开发新型绿色抑菌剂奠定了基础。 |
| 关键词: 奶牛乳房炎 金黄色葡萄球菌噬菌体 裂解酶 克隆 蛋白结构 |
| DOI： |
| 投稿时间：2016-08-26修订日期：2016-09-28 |
| 基金项目:国家自然科学基金项目（31560699），新疆生产建设兵团应用基础研究计划项目(2015AG012） |
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| Cloning and sequence analysis of bovine mastitis Staphylococcus aureus lytic bacteriophage lysin LysIMEP5 gene |
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| (College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang) |
| Abstract: |
| Cloning and analysis the biological information of lysin LysIMEP5 gene from bovine mastitis Staphylococcus aureus lytic bacteriophage. Using the bovine mastitis Staphylococcus aureus lytic phage VB_SauS_IMEP5, which was isolated and indentified by our laboratory, and its whole genome information, analysis, we got the lysin gene sequence. Useing the software Primer5.0, a pair of specific primers was designed and the LysIMEP5 gene was amplified and cloned from the bovine mastitis Staphylococcus aureus phage by Polymerase Chain Reaction (PCR). Analysing its biological information characteristics by the BLAST and its protein structure prediction. The lysin LysIMEP5 gene was cloned successfully and its sequencing results were completely matched with the original sequence by DNAMAN, without any gene mutation. Homologous analysis showed that,the highest similarty is 83.1% compared with have reported Staphylococcus aureus phage lyase.The gene full-length of lysin LysIMEP5 was 1371bp, encoding 456 amino acids. The lysin LysIMEP5 was hydrophilic protein, its molecular mass was 51.717 kDa, its isoelectric point was 9.70. Biological information analysis showed that, CHAP fragments and Amidase-3 fragments were existed in the LysIMEP5 at the same time. The protein was mainly random coil, but without transmembrane domain and signal peptide. The gene of LysIMEP5 was successfully cloned from the bovine mastitis Staphylococcus aureus phage and the protein structure of LysIMEP5 was prediction. The results provided foundation for the future expression of LysIMEP5 and the development of new green antibacterial agent. |
| Key words: bovine mastitis Staphylococcus aureus bacteriophage lysine cloning protein structure |
