摘要
根据GenBank中的lmo2193基因序列设计特异性引物,利用PCR方法对lmo2193基因进行扩增,将扩增产物克隆到pMD19-T载体,测序后对基因核苷酸序列进行分析,预测其编码蛋白质的二三级结构,对其进行同源性及遗传变异分析。新疆分离株LM90SB2的lmo2193基因序列全长为1303 bp,包含1077 bp开放阅读框,共编码358个氨基酸;LM90SB2株lmo2193基因核苷酸序列与NTSN、F2365、81-0582、10-0809、02-1792相似性为99.99%;与SLCC2755、H34、WSLC1019相似性为99.3%~99.7%;与C1-387、L2625、10-4754、lm3136相似性为98.2%~98.3%。推导的氨基酸同源性为95.3%~99.7%。分子进化树表明,LM90SB2菌株lmo2193基因与4b型菌株的亲缘关系较近。蛋白质二级结构预测表明,LM90SB2 lmo2193蛋白为亲水性蛋白,无信号肽,不形成跨膜结构。蛋白结构域预测,lmo2193蛋白为ATP酶。成功克隆LM90SB2基因,为进一步研究LM90SB2的lmo2193基因功能提供一定依据。
Specific primers were designed according to the lmo2193 gene sequence in GenBank.The PCR method was used to amplify the lmo2193 gene.The amplified product was cloned into the pMD19-T vector,and the nucleotide sequence of the gene was sequenced and analysized.We predicted the two-level structure of the encoded protein and analyzed its homology and the genetic variation.The sequence length of LM90SB2 lmo2193 gene was 1303 bp,containing a 1077 bp open reading frame encoding 358 amino acids.The nucleotide sequence similarity of lmo2193 gene of LM90SB2 strain was 99.9%with NTSN,F2365,81-0582,10-0809,02-1792;99.3%to 99.7%with SLCC2755,H34,WSLC1019;and 98.2%to 98.3%with C1-387,L2625,10-4754,lm3136.The putative amino acid sequence similarity was 95.3%to 99.7%with above strains.The phylogenetic tree shows that the lmo2193 gene of LM90SB2 strain was closely related to the serotype 4 b strain.The protein secondary structure prediction showed that LM90SB2 lmo2193 protein was a hydrophilic protein without signal peptide and transmembrane structure.The protein domain predicted that the lmo2193 protein was an ATPase.The lmo2193 gene of LM90SB2 strain was successfully cloned and provided a theretical basis for further study of lmo2193 gene.
作者
李红欢
马勋
钱凌霄
杜冬冬
张奇文
李庆辉
刘扬扬
Li Honghuan;Ma Xun;Qian Lingxiao;Du Dongdong;Zhang Qiwen;Li Qinghui;Liu Yangyang(College of Animal Science and Technology,Shihezi University,Shihezi,832000)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2019年第11期5017-5022,共6页
Genomics and Applied Biology
基金
国家自然基金项目(31360614)资助
