奶牛源葡萄球菌tetK基因碱基及氨基酸序列差异的功能分析

发布时间：2018-06-13 15:55

本文选题：奶牛源葡萄球菌 + 四环素耐药基因tetK　；参考：《扬州大学学报(农业与生命科学版)》2017年02期

【摘要】：为探究奶牛源葡萄球菌中部分四环素耐药基因tetK不产生耐药性的原因,采用平行试验对四环素抗性菌株(MP00159)与四环素敏感菌株(MP00109)所携带的tetK基因和tetK′基因的碱基及氨基酸序列差异进行分析;随后将两者分别克隆至表达载体pET-32a(+)后导入大肠埃希菌感受态细胞BL21(DE3)内,比较转化前后对四环素的耐药性。结果表明:四环素敏感菌株携带的tetK′基因与抗性菌株的tetK基因存在5个碱基位点差异,对应的氨基酸出现4个差异;转入敏感菌株tetK′基因的BL21仍表现对四环素敏感,转入抗性菌株tetK基因的BL21表现了四环素抗性。由此推测发生突变的碱基位点是决定tetK耐药表型的关键位点。
[Abstract]:In order to investigate the reasons why tetk, a tetracycline resistant gene, was not resistant to antibiotics in staphylococcus from dairy cattle, The nucleotide and amino acid sequence differences of tetK gene and tetK' gene carried by tetracycline resistant strain (MP00159) and tetracycline sensitive strain (MP00109) were analyzed by parallel test. Then they were cloned into the expression vector pET-32a () and introduced into Escherichia coli competent cell BL21 (DE3) to compare the resistance of tetracycline to tetracycline before and after transformation. The results showed that the tetK' gene carried by tetracycline sensitive strain was different from that of resistant strain in 5 base loci, and the corresponding amino acid was 4 different, while BL21, which was transferred to tetK' gene, was still tetracycline sensitive. BL21, which was transformed into resistant strain tetK, showed tetracycline resistance. Therefore, it is inferred that the base site of mutation is the key site to determine tetK resistance phenotype.
【作者单位】：扬州大学兽医学院/教育部禽类预防医学重点实验室/江苏省动物重要疫病与人兽共患病防控协同创新中心;扬州大学实验农牧场;常州市武进区动物疫病预防控制中心;
【基金】：江苏省青蓝工程项目(2012-05) 扬州市社会发展科技攻关项目(YZ2011091) 江苏省农业科技自主创新资金项目[CX(14)2088] 江苏高校优势学科建设工程项目(PAPD) 常州市科技支撑计划项目(CE20152006)
【分类号】：S852.611

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