单增李斯特菌llsB基因克隆及序列分析

doi:10.16431/j.cnki.1671-7236.2019.07.002

Abstract

Abstract:

The aim of this study was to clone and analyze llsB gene of Listeria monocytogenes LM90SB2 isolated from encephalitis sick sheep in Xinjiang,and further improve the functional study of listeriolysion S.Specific primers were designed according to the full-length sequence of Listeria monocytogenes F2365 gene in GenBank (accession No.:AE017262),llsB gene of Xinjiang isolate LM90SB2 was amplified by PCR,and recycled the target gene and connected with pMD19-T vector.Positive bacteria was identified and screened for sequencing by PCR and double enzyme digestion methods.The nucleotide sequence with the correct sequencing result was analyzed,and the homology and genetic variation analysis were carried out.The results showed that llsB gene of Xinjiang isolate LM90SB2 was 876 bp in length and encoded a total of 291 amino acids.The nucleotide homology of llsB gene were 100% between LM90SB2 isolate and 10-0809,81-0592,81-0558,02-1792,NTSN and 02-1289 isolate strains,respectively,the homology with CⅡMS-PH-1 and NRRLB-57603 were both 99.9%,and the homology with J1816 and R2-502 were 44.7% to 45.0%.The molecular phylogenetic tree showed that llsB gene of LM90SB2 strain was closely related to the strain with serotype 4b,and the clustering was the same branch.Protein secondary structure prediction results showed that LM90SB2 llsB protein was a hydrophilic protein,without signal peptide and transmembrane structure.This experiment successfully cloned LM90SB2 llsB gene,and provided a comprehensive theoretical basis for the function research of LM90SB2 llsB gene.

Key words: Listeria monocytogenes; llsB gene; cloning; bioinformatics analysis

CLC Number:

S852.61

WANG Xinyu, YIN Hua, RENG Jingjing, LI Honghuan, JIANG Jianjun. Cloning and Sequence Analysis of Listeria monocytogenes llsB Gene[J]. , 2019, 46(7): 1890-1898.

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Cloning and Sequence Analysis of Listeria monocytogenes llsB Gene

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