摘要
单核细胞增多性李斯特菌(Listeria monocytogenes,LM)作为四大重要食源性病原菌之一,对公共安全及畜牧业发展威胁极大.其lismff_0038是新发现的一个潜在毒力因子,与LM的神经感染和母胎感染密切相关.对食源性单核细胞增多性李斯特菌LM5567的lismff_0038基因进行克隆和生物信息学分析,为功能验证提供基础.根据GenBank中收录的lismff_0038基因序列设计特异性引物,利用PCR方法对食源性单核细胞增多性李斯特菌LM5567的lismff_0038基因进行扩增,将扩增产物克隆到pMD19-T载体,进行PCR、双酶切鉴定及序列测定,并对基因核苷酸序列和蛋白序列进行分析.结果显示,食源性单核细胞增多性李斯特菌LM5567的lismff_0038基因序列全长为988 bp.LM5567的lismff_0038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株(4b,奶酪,加拿大)、81-0861株(4b,卷心菜,加拿大)相似性91.1%.其推导的氨基酸序列与上述菌株相似性100%.蛋白质二级结构预测表明,LM5567的lismff_0038蛋白为疏水性蛋白,无信号肽,属于跨膜蛋白.表明,克隆了LM5567的lismff_0038基因,为进一步探究lismff_0038基因的功能奠定了一定基础.
Listeria monocytogenes(LM),one of the four important foodborne pathogens,poses a great threat to public safety and animal husbandry.Its lismff_0038 is a newly discovered potential virulence factor,which is closely related to LM nerve infection and maternal infection.Cloning and bioinformatics analysis of the lismff_0038 gene of Listeria monocytogenes LM5567 provides a basis for functional verification.The specific primers were designed according to the lismff_0038 gene sequence contained in GenBank.The lismff_0038 gene of Listeria monocytogenes LM5567 was amplified by PCR and the amplified product was cloned into pMD19-T vector.PCR,double enzyme digestion and sequencing were performed,and the nucleotide sequence and protein sequence of the gene were analyzed.The results showed that,the full length of the lismff_0038 gene of Listeria monocytogenes LM5567 was 988 bp.LM5567 lismff_0038 nucleotide sequence with 02-6680 strain(4b,cheese,Canada),10-0811 strain(1/2b,crab,Canada),10-0809 strain(4b,feces,Canada),81-0558 strain(4b,cerebrospinal fluid,Canada),02-1103 strain,02-1792(4b,cheese,Canada),81-0861 strain(4b,cabbage,Canada)similarity 91.1%.The deduced amino acid sequence is 100%similar to the above strain.Protein secondary structure prediction indicated that the lismff_0038 protein of LM5567 is a hydrophobic protein,no signal peptide,and belongs to the transmembrane protein.Conclusion The lismff_0038 gene of LM5567 was cloned,which laid a foundation for further exploration of the function of lismff_0038 gene.
作者
刘扬扬
马勋
康立超
李红欢
钱凌霄
江婉琳
阮婷玉
Liu Yangyang;Ma Xun;Kang Lichao;Li Honghuan;Qian Lingxiao;Jiang Wanlin;Ruan Tingyu(College of Animal Science and Technology,Shihezi University,Xinjiang Shihezi 832003;Analysis and Testing Center,Xinjiang Academy of Agricultural and Reclamation Science,Xinjiang Shihezi 832000)
出处
《现代畜牧兽医》
2019年第10期1-7,共7页
Modern Journal of Animal Husbandry and Veterinary Medicine
基金
国家自然科学基金项目（31860712）
