奶牛腐蹄病坏死杆菌分离鉴定及白细胞毒素基因lktA1序列分析

doi:10.16431/j.cnki.1671-7236.2015.09.014

›› 2015, Vol. 42 ›› Issue (9): 2308-2312.doi: 10.16431/j.cnki.1671-7236.2015.09.014

Isolation,Identification and Sequence Analysis of Leukotoxin Gene lktA1 of Fusobacterium necrophorum of Cow Footrot

GAO Jing, LI Chun-qiu, YAO Shuang, ZHANG Li-chun, WANG Xin-yu, WANG Zhi-hui, ZHAO Xi-wen, WEI Shan, WANG En-yu, SUN Dong-bo, WU Rui, GUO Dong-hua

College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing 163319, China

Received:2015-04-14 Online:2015-09-20 Published:2015-09-25

Abstract

Abstract: For separation and purification of Fusobacterium necrophorum of cow footrot, and analysis of genetic relationship with other strains, the hoof ministry swab samples were detected by PCR based on specific primers of leukotoxin gene, and genomic DNA were isolated from PCR positive samples of Fusobacterium necrophorum culturing in anaerobic medium.The genes of leukotoxin were cloned and sequenced.The results showed that nine of hoof ministry swab samples were all PCR positive samples, and we obtained Fusobacterium necrophorum pure culture from one of the samples which named bFR13-1.The gene sequencing results indicated that the homologies of leukotoxin gene nucleotide sequence of bFR13-1 strain compared with H05, A25 and B35 strains from GenBank were 98.40%, 98.35% and 90.79%, respectively, and the homologies of deduced amino acid sequence were 97.7%, 97.6% and 89.0%, respectively.Phylogenetic tree analysis results showed that leukotoxin gene of Fusobacterium necrophorum bFR13-1 and H05 had high homology and bFR13-1, H05 and A25 showed a close genetic relationship.The result indicated that leukotoxin showed variability between different Fusobacterium necrophorum isolated strains, and it was worth to study whether this change and pathogenicity of Fusobacterium necrophorum were related.

Key words: cow footrot; Fusobacterium necrophorum; leukotoxin

CLC Number:

S852.61

GAO Jing, LI Chun-qiu, YAO Shuang, ZHANG Li-chun, WANG Xin-yu, WANG Zhi-hui, ZHAO Xi-wen, WEI Shan, WANG En-yu, SUN Dong-bo, WU Rui, GUO Dong-hua. Isolation,Identification and Sequence Analysis of Leukotoxin Gene lktA1 of Fusobacterium necrophorum of Cow Footrot[J]. , 2015, 42(9): 2308-2312.

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Isolation,Identification and Sequence Analysis of Leukotoxin Gene lktA1 of Fusobacterium necrophorum of Cow Footrot

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