水貂IgG Fc段基因及抗原性分析

GENETIC AND ANTIGENIC ANALYSIS OF FC REGION OF MINK IGG

根据GenBank上发布的水貂IgG中Fc段的序列(L07789.1),设计合成1对引物用于扩增水貂Fc段基因。从水貂脾脏中提取总RNA,RT-PCR扩增水貂Fc段基因,获得大小约606 bp的片段,将其连接T载体并测序。然后进行遗传进化分析,在此基础上利用Western blot进行抗体杂交试验。结果显示水貂IgG Fc与Fc犬核苷酸序列同源性为85.0%,氨基酸序列同源性为80.5%。Western blot显示水貂IgG重链可以和抗犬IgG有效结合,遗传进化分析得出哺乳动物中水貂与犬IgG Fc的亲缘关系最近。通过以上分析和相关文献得出用犬的诊断试剂可以检测水貂疾病,或用犬的抗血清进行水貂疾病治疗。

Detection methods for distemper and canine parvovirus can be universal for dogs and minks according to cloning and phylogenetic analysis of Fc gene as well as reactivity of mink IgG in Western blot. In order to obtain a certain length of Fc segment gene, a pair of primers were designed based on Fc gene sequences of mink IgG (L07789) published in GenBank. Total RNA materials were extracted from spleens of minks and amplified in RT-PCR. The results showed that the Fc segment gene consisted of 606 bp. Amplified Fc fragment was then cloned into the vector PEasy-T1 and sequenced for phylogenetic analysis. Antibody hybridization test was performed in Western blot. The nucleotide sequence homology between mink and canine was 85.0%and amino acid sequence homology was 80.5%. The heavy chain of mink IgG reacted with anti-dog IgG in Western blot. It was concluded that the Fc gene of mink and dog IgGs had the closest relationship within mammalians based on genetic evolution. The results obtained from the preceding study and related literature confirmed that dog and mink diseases could be mutually diagnosed and treated using the reagents of either dog or mink origin.