鸭疫里默氏杆菌中国分离株的外膜蛋白A基因的克隆与序列分析

为了对鸭疫里默氏杆菌(Riemerella anatipestifer,RA)中国分离株的外膜蛋白A(OmpA)基因的序列进行分析,根据GenBank中已发表的鸭疫里默氏杆菌(Riemerella anatipestifer,RA)的外膜蛋白A(Om-pA)基因序列,在其高度保守区设计一对特异性引物,应用PCR技术,分别扩增从中国广东、福建和山东等地分离的8株鸭疫里默氏杆菌中国分离株的OmpA基因,克隆测序后与GenBank中已发表的26株鸭疫里默氏杆菌进行序列比较,结果表明:所有菌株的OmpA基因均为由1164个碱基组成的ORF,编码387个氨基酸组成的蛋白质,起始密码子均为ATG,终止密码子均为TAA,其核苷酸序列非常保守,同源性高达88.2%~100%,氨基酸同源性达到88.9%~100%。试验表明,OmpA基因具有种内相对保守性,其核苷酸与氨基酸同源性与鸭疫里默氏杆菌的血清型、分离时间和地点之间无必然联系。

Cloning and Sequencing of Outer Membrane Protein A of Chinese Riemerella anatipestifer

In order to analyze sequences of OmpA of Chinese strains of Riernerella anatipestifer(RA), according to outer membrane protein A(OmpA) gene sequences from R.anatipestifer strains in the GenBank, apair of specific primers was designed.Eight R.anatip estif er strains from Fujian, Guanglong and Shandong were amplified in the whole open reading frame(ORF) by PCR and then cloned and sequenced.The comparative analysis of the sequences with twenty-six R.anatip estifer strains in GenBank indicated all OmpA gene had a long complete ORF composed of 1164 nucleotides which encoled a protein of 387 amino acids, and all initiation colon were ATG and end codon were TAA.The sequence of OmpA were relatively conservative which identity was 88.2%~100% and the identity of the amino acids was 88.95%~100%.The results showed that the sequence of OmpA relatively conservative and the identity of the nucleotide and amino acid had no positive relation with the isolate serotype, the time and location of isolation.