16S rRNA序列分析在猪肠道细菌鉴定中的应用

Application of Sequence Analysis of 16S rRNA Gene in Identifying Porcine Intestinal Bacteria

为建立16SrRNA基因序列分析鉴定猪肠道细菌的方法,选择细菌的16SrRNA基因为靶序列设计引物,对11株猪肠道细菌进行PCR扩增,并对PCR产物进行测序和序列分析。序列分析结果显示,A2、A4序列与柠檬明串珠菌同源性>99%;B2序列与鼠乳杆菌同源性>99%;C2序列与唾液乳酸杆菌同源性>99%;F3序列与类肠膜魏斯氏菌同源性>97%;F6和F7序列与融合魏斯氏菌同源性>99%;M3和M8序列与洛菲氏不动杆菌同源性>98%;M9和M11序列与粪肠球菌同源性>99%。表明建立的16SrRNA基因序列分析方法可以应用于鉴定猪肠道细菌。

To develop a technique for bacterial molecular identification and evaluate its effect on identification of porcine intestinal bacteria,a serial of primers targeting 16S rRNA genes were chosen to amplify certain fragments by PCR.After sequencing,the nucleotide sequences were compared with those previously reported in GenBank database.The BLAST analysis and phylogenetic analysis indicated that the 16S rRNA genes of A2 and A4 were most similar to that of Leuconostoc citreum（similarity99%）,while B2 was most similar to Lactobacillus murinus（similarity99%）.For C2,F3,F6 and F7,the 16S rRNA genes were most similar to those of Lactobacillus salivarius（similarity99%）,Weissella paramesenteroides（similarity97%）,and Weissella confusa（similarity99%）,respectively.The 16S rRNA genes of M3 and M8 were most similar to that of Acinetobacter lwoffii（similarity98%）,while for M9 and M11,they were most similar to Enterococcus faecalis（similarity99%）.This indicates that the 16S rRNA gene sequence technology is feasible for identifying porcine intestinal bacteria.