摘要
通过提取猪传染性胸膜肺炎放线杆菌ApxⅠA基因组DNA,运用PCR方法扩增ApxⅠA基因片段,并采用TA克隆技术获得pMD18–T–ApxⅠA重组质粒,进行生物信息学分析。结果表明:成功获得长度为1 209 bp的ApxⅠA基因;ApxⅠA蛋白的理化性质稳定;ApxⅠA蛋白有磷酸化位点,但无糖基化位点;信号肽预测ApxⅠA蛋白无信号肽;ApxⅠA蛋白为亲水性蛋白,无跨膜结构;预测二级结构主要由无规则卷曲构成,占50.62%,α–螺旋占25.56%,β–折叠占23.82%,三级结构主要由无规则卷曲和β–折叠构成。
The genomic DNA was extracted from porcine Actinobacillus pleuropneumoniae and subjected to PCR using ApxⅠA gene primers. The resulted product was linked to pMD18–T vector and then generated pMD18–T–ApxⅠA recombinant plasmid for bioinformatics analysis. The biochemical results indicated that 1 209 bp target fragment of ApxⅠA was managed to be cloned. And bioinformatics analysis revealed that ApxⅠA protein had a stable physicochemical property and had a phosphorylation site and no glycosylation site. The online signal peptide prediction showed that ApxⅠA protein was hydrophilic protein without trans–membrane domains and signal peptide. Structure prediction revealed its tertiary structure consisted of 25.56% of α–helixes, 23.82% of β–sheets and 50.62% of radon coils.
作者
阮业召
姜家麟
罗维
郭小权
胡国良
刘平
RUAN Yezhao;JIANG Jialin;LUO Wei;GUO Xiaoquan;HU Guoliang;LIU Ping(Institute of Animal Disease Surveillance and Prevention,Jiangxi Agricultural University,Nanchang,Jiangxi 330045,China;Hunan SJA Laboratory Animal Co.Ltd.,Changsha,Hunan 410125,China)
出处
《湖南农业大学学报（自然科学版）》
CAS
CSCD
北大核心
2018年第5期544-548,共5页
Journal of Hunan Agricultural University(Natural Sciences)
基金
国家自然科学基金项目(31402266)
江西省科技支撑计划项目(20141BBF60035)
江西省自然(青年)科学基金项目重点项目(20171ACB21026)
江西省教育厅科学技术研究重点项目(GJJ170243)
江西省研究生创新专项资金项目(YC2017–S185)
关键词
猪传染性胸膜肺炎
放线杆菌
ApxⅠA基因
分子克隆
生物信息学分析
porcine contagious pleuropneumonia（PCP）
Actinobacillus pleuropneumoniae
ApxⅠA gene
molecular cloning
bioinformatics analysis
