白来航鸡MHC-Ⅰ重链基因克隆与序列分析

Cloning and sequencing of Gallus chicken MHC-Ⅰ heavy chain gene

参照GenBank发表的鸡MHC-Ⅰ重链（BF2）基因序列,设计并合成1对引物,无菌采取纯系来航SPF鸡抗凝静脉血,直接提取总RNA,采用RT-PCR技术,PCR扩增BF2基因,并进行克隆和序列测定。测序结果表明,获得了来航鸡BF2基因,其大小为1 035 bp,包含一个完整阅读框,共编码345个氨基酸。与GenBank上鸡BF2基因序列进行比较,其核苷酸同源性在93.0%~97.6%之间,氨基酸同源性在83.2%~97.1%之间。来航鸡与人和其他种动物的MHC-Ⅰ重链基因核苷酸同源性在12.9%~83.6%之间。

One pair of primers was designed according to chicken MHC-Ⅰ heavy chain（BF2） gene sequences published in GenBank.Chicken BF2 gene was amplified by RT-PCR from the total RNA of peripheral blood mononuclear cells（PBMCs） isolated from Gallus SPF chicken,and cloned into pGEM-T easy vector.The result showed that the full length of chicken BF2 gene is consisted of 1035 bp,which includes an open reading frame（1035 bp）,encoding 345 amino acid residues.The nucleotide sequence of homology to the chicken MHC-Ⅰ heavy chain gene was between 93.0% and 97.6%.The deduced amino acids sequence of homologues to those of the chicken MHC-Ⅰ heavy chain gene were between 83.2% and 97.1%.Comparison of the nucleotides sequence of chicken MHC-Ⅰ heavy chain with those from human and other animals showed that the nucleotides homologues were between 12.9% and 83.6%.