16S rRNA序列分析在皖南花猪病原菌分离鉴定中的应用研究
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摘要
为了解地方品种皖南花猪细菌感染情况,运用16S rRNA基因序列分析方法鉴定从皖南花猪地方品种猪分离的8种细菌.通过提取细菌DNA,采用通用引物PCR扩增16S rRNA基因片段并测序.将测序结果用BLAST在线软件在Nucleotide数据库中进行序列同源性比对,根据序列同源性鉴定病原细菌.结果表明:7种细菌鉴定到"种",1种细菌鉴定到"属". 16S rRNA基因序列分析对于皖南花猪病原菌鉴定是一种有效的方法 .
In order to understand the bacterial infection of the local pig breed in southern Anhui,8 kinds of bacteria isolated from the local pig breed in southern Anhui were identified by sequencing the 16 SrRNA gene. DNA samples of the 8 bacteria were extracted. The 16 SrRNA genes were amplified by PCR and sequenced with universal primers. The sequences of the 16 SrRNA genes were aligned by online software BLAST in nucleotide database. The bacteria were identified according to the homology of their 16 SrRNA genes. The results show that seven kinds of bacteria were classified as species; another bacterium was identified as genus. 16 SrRNA gene sequence analysis is useful in identifying pathogenic bacteria of the Pig in Southern Anhui.
In order to understand the bacterial infection of the local pig breed in southern Anhui,8 kinds of bacteria isolated from the local pig breed in southern Anhui were identified by sequencing the 16 SrRNA gene. DNA samples of the 8 bacteria were extracted. The 16 SrRNA genes were amplified by PCR and sequenced with universal primers. The sequences of the 16 SrRNA genes were aligned by online software BLAST in nucleotide database. The bacteria were identified according to the homology of their 16 SrRNA genes. The results show that seven kinds of bacteria were classified as species; another bacterium was identified as genus. 16 SrRNA gene sequence analysis is useful in identifying pathogenic bacteria of the Pig in Southern Anhui.
引文
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[7]Drancourt M,Bollet C,Carlioz A,et al.16S ribosomal DNA sequenceanalysis of a large collection of environmental and clinical unidentifiablebacterial isolates[J].J Clin Microbiol,2000,38(10):3623-3630.
[8]Kimura M.A simple method for estimating evolutionary rates of basesubstitutions through comparative studies of nucleotide sequences[J].JMol Evol,1980,16(2):111-120.
[9]Thorne J L,Kishino H,Painter I S.Estimating the rate of evolution of therate of molecular evolution[J].Mol Biol Evol,1998,15(12):1647-1657.
[10]张玲玲.16S rRNA序列分析法在医学微生物鉴定中的应用研究[J].医药化工,2017,43(12):178-179.
[11]邱吉宇,主性,钟纯燕,等.贵州中蜂蜂房哈夫尼菌的分离鉴定及药敏试验[J].中国畜牧兽医,2018,45(4):1050-1058.
[12]赵全邦,胡广卫,李静,等.青海血蜱和森林革蜱的分子生物学鉴定[C]//中国动物学会寄生虫学专业委员会会议论文集,2018:126.
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