3株不同毒力巴氏杆菌外膜蛋白H基因克隆及序列分析

Cloning and Sequencing of Outer Membrane Protein H Gene of 3 Different Pasteurella multocide Speices with Different Virulence

参照GenBank中巴氏杆菌外膜蛋白H（ompH）基因核苷酸序列，设计了一对特异性引物，通过PCR方法，扩增了3株不同毒力猪源巴氏杆菌CA4—1株、679—230株、3397A株ompH全基因。PCR产物克隆至pMD-18T载体，进行序列分析，发现3个菌株ompH基因完整的ORF大小分别为1008bp、1008bp和1047bp，分别编码336、336、349个氨基酸，它们的前20个氨基酸均组成信号肽。与已知的15个Heddleston血清型的核苷酸及其推导的氨基酸序列比较，核苷酸同源性在83．3％-99．8％，氨基酸同源性在79．2％-99．4％。序列分析结果表明，ompH基因在不同血清型之间具有较高的同源性。

According to the sequence of ompH of Pasteurella multocida published in GenBank, a set of specific primers were designed. Using the primers, full - length ompH gene of 3 different Pasteurella multocide speices（ CA4 -1,679 -230, 3397A） with different virulence was amplified by PCR. The PCR products were cloned into pMD18 - T vector for sequencing. Sequence analysis showed that the complete ORF of ompH gene of 3 species were 1008bp, 1008bp and 1047bp, encoding 336, 336,349 amino acid respectively. Each predicted primary protein has a 20 - amino - acid signal peptide in amino terminal. Compared with ompH gene from 15 Heddleston serotypes,it showed that nucleotide sequence homologies were 83.3% to 99.8%, and the deduced amino acid homologies were 79.2% to 99.4%. The sequence analysis result indicated that ompH is of hign homology between different serotypes.