猪葡萄球菌EXHC、SHETA基因的克隆、序列分析及蛋白结构预测  

Cloning and Sequence Analysis of EXHC and SHETA Genes of Staphylococcus hyicus and Prediction of Protein Structure

作　　者：张乐宜[1] 李艳[1] 蔡汝健[1] 蒋智勇[1] 刘燕玲[1] 宋长绪[1] 

机构地区：[1]广东省农业科学院动物卫生研究所、广东省兽医公共卫生公共实验室,广东广州510640

出　　处：《中国畜牧兽医》2014年第10期17-22,共6页China Animal Husbandry & Veterinary Medicine

基　　金：广东省科技厅项目(2012A020200014、2010A04030-1010);广州市科技计划项目(12C14071615);广东省生猪产业体系项目(粤农2009-380);广东省农业科学院院长基金(201313)

摘　　要：为分析猪葡萄球菌脱落毒素EXHC和SHETA基因结构并预测其所编码蛋白的结构和功能,本试验根据GenBank已报道的猪葡萄球菌脱落毒素EXHC和SHETA基因序列分别设计了1对特异性引物,从猪葡萄球菌GDZC株中扩增获得EXHC和SHETA基因片段,大小分别为1007和971bp。序列分析结果表明,EXHC基因与猪葡萄球菌丹麦分离株（GenBank登录号：AF515455）及猪源松鼠葡萄球菌河北分离株（GenBank登录号：JF755400）的核苷酸序列、氨基酸序列同源性均为100.0%,而与其他葡萄球菌脱落毒素基因的核苷酸、氨基酸序列同源性分别为3.3%~53.9%和7.9%~44.2%。SHETA基因与日本分离株（GenBank登录号：AB036768）的核苷酸序列同源性为96.2%,氨基酸序列同源性为98.4%,在保守区共有31个碱基发生突变。利用DNAStar软件对EXHC和SHETA蛋白结构进行分析,结果显示EXHC基因编码的蛋白为亲水性蛋白,SHETA基因编码蛋白为疏水性蛋白,抗原性较差。本研究从猪葡萄球菌GDZC株中成功扩增获得EXHC和SHETA基因片段并对其编码的蛋白结构进行了预测,证实了中国分离的猪葡萄球菌同时携带有EXHC和SHETA2种毒素基因,为进一步研究猪葡萄球菌的致病机理及毒素之间的相互作用提供了理论依据。It was to analyze and predict the structural characteristics of EXHCand SHETAgenes fromStaphylococcus hyicus exfoliative toxins by bioinformatics.According to Staphylococcus hyicus exfoliative toxins EXHCand SHETAgenes sequence reported in GenBank,two pairs of primers were designed to amplify EXHCand SHETAgene fragments,respectively.The fragment size of EXHCand SHETAgenes of GDZC strains were 1007 and 971bp,respectively.The nucleotide and amino acid sequences analysis showed that EXHCgene exhibited homology 100.0% with Staphylococcus hyicus Denmark strain（AF515455）and Staphylococcus sciuri Hebei strain（JF755400）,while shared low homology,3.3%to 53.9% and 7.9%to44.2% with exfoliative toxin of other Staphylococci stains based on nucleotide and amino acid sequence,respectively.There were 31 nucleotide mutations in the conserved region of SHETAgene,the nucleotide and amino acid sequence homologies with the Japan isolates（AB036768）were 96.2%and 98.4%,respectively.Using the DNAStar software to analyze the structure of proteins ecoded by EXHCand SHETAgenes,the results showed that protein encoded by gene EXHC was hydrophilic protein,and protein encoded by gene SHETAwas hydrophobic and poor antigenicity.This study amplified the EXHCand SHETAgenes fromStaphylococcus hyicus GDZC strains and predicted the protein structure successfully,and confirmed Staphylococcus hyicus from China could carry two toxins genes EXHCand SHETA,provided a theoretical basis for further study of the Staphylococcus hyicus pathogenic mechanism and interaction between toxins.

关 键 词：猪葡萄球菌 表皮脱落毒素 EXHC SHETA 

分 类 号：S858.28[农业科学—临床兽医学]