食源性单核细胞增生李斯特菌lmoF2365_0037基因克隆及序列分析

Cloning and sequence analysis of lmo F2365_0037 in foodborne Listeria monocytogenes

为了对食源性单核细胞增生李斯特菌LM5567的lmoF23650037基因进行克隆和序列分析。试验利用PCR方法对lmoF23650037基因进行扩增,连接pMD19-T载体进行克隆,筛选阳性菌株进行测序比对分析。结果显示:扩增获得的lmoF23650037基因序列长为548 bp,克隆得到lmoF23650037基因的阳性转化子;生物信息学分析表明:lmoF23650037蛋白属于跨膜蛋白,无信号肽序列;二级结构中无规则卷曲和延伸链比例较大,分别是46.31%和32.89%;序列比对结果表明,LM5567株lmoF23650037基因核苷酸序列与02-6680株(4b,奶酪,加拿大)、10-0811株(1/2b,螃蟹,加拿大)、10-0809株(4b,粪便,加拿大)、81-0558株(4b,脑脊髓液、加拿大)、02-1103株、02-1792(4b,奶酪,加拿大)、81-0861株(4b,卷心菜,加拿大)相似性均为100%;LM5567 lmoF23650037基因编码的氨基酸序列与上述菌株相似性均为93.3%。本试验成功克隆了lmoF23650037基因,为进一步探究lmoF23650037基因的功能奠定了基础。

For cloning and sequence analysis of the lmoF23650037 gene of Listeria monocytogenes Listeria LM5567.The lmoF23650037 gene was amplified by PCR and ligated with pMD19-T vector.The positive strains were screened for sequencing analysis.The results showed that the amplified lmoF23650037 gene sequence was 548 bp long,and the positive transformant of lmoF23650037 gene was cloned.Bioinformatics analysis showed that lmoF23650037 protein belongs to transmembrane protein,no signal peptide sequence;no regular curl and extension in secondary structure The proportion of the chain was 46.31%and 32.89%,respectively;the sequence alignment showed that the nucleotide sequence of LM5567 strain lmoF23650037 gene and 02-6680 strain(4 b,cheese,Canada),10-0811 strain(1/2 b,crab),Canada),10-0809 strains(4 b,feces,Canada),81-0558 strains(4 b,cerebrospinal fluid,Canada),02-1103 strains,02-1792(4 b,cheese,Canada),81-0861 strains The similarity of(4 b,cabbage,Canada)was 100%;the amino acid sequence encoded by LM5567 lmoF23650037 gene was 93.3%similar to the above strain.The lmoF23650037 gene was successfully cloned in this experiment,which laid a foundation for further exploration of the function of lmoF23650037 gene.