朝鲜鹌鹑MHC class Ⅰ基因第2、3外显子多态性与NDV抗性的关联分析  

Association of polymorphism in exons 2 and 3 of MHC class Ⅰ gene of Korean quail and Newcastle disease virus resistance

作　　者：崔小辉[1] 赵淑娟[1] 张小辉[1] 庞有志[1] 杨又兵[1] 白俊艳[1] 雷莹 CUI Xiaohui;ZHAO Shujuan;ZHANG Xiaohui;PANG Youzhi;YANG Youbing;BAI Junyan;LEI Ying(College of Animal Science and Technology , Henan University of Science and Technology Luoyang, Henan 471003 , China)

机构地区：[1]河南科技大学动物科技学院,河南洛阳471003

出　　处：《西北农林科技大学学报（自然科学版）》2019年第6期1-6,共6页Journal of Northwest A&F University(Natural Science Edition)

基　　金：河南省产学研合作项目(2015HNCXY012);河南科技大学高级别项目培育基金项目(2015GJB028)

摘　　要：【目的】研究朝鲜鹌鹑MHC classⅠ基因第1,2和3外显子的多态性,并与新城疫病毒抗性进行关联分析。【方法】采用胸肌注射的方法对14日龄朝鲜鹌鹑进行新城疫La Sota弱毒株攻毒,每周注射1次,连续攻毒3周后,心脏采血,采用β-微量法测定血清中新城疫抗体水平。PCR扩增朝鲜鹌鹑MHC classⅠ基因,测序后采用DNA Star、SPSS等分析软件研究朝鲜鹌鹑MHC classⅠ基因第1,2,3外显子的多态性,并分析不同基因型新城疫抗体水平的差异。【结果】朝鲜鹌鹑的新城疫抗体效价为26～210。PCR扩增获得了1 290bp的MHC classⅠ基因,其编码区序列为780bp,编码259个氨基酸。通过与已知序列日本鹌鹑RNA(AB005528)比对,得到长度分别为76,272和274bp的第1,2和3外显子。在第1外显子上未检测到突变为点,在第2和3外显子上共检测到11个突变位点(第2外显子上4个突变位点:257bp(A/C)、267bp(A/T)、366bp(A/G)和500bp(T/G);第3外显子上7个突变位点:795bp(T/C)、798bp(T/C)、810bp(T/G)、830bp(A/G)、873bp(A/T)、927bp(A/G)和932bp(C/G)),均为中度多态基因座,11个突变位点中,267bp(A/T)、830bp(A/G)、873bp(A/T)、932bp(C/G)突变导致编码氨基酸改变,500bp(T/G)、873bp(A/T)、932bp(C/G)突变位点基因型之间新城疫抗体水平存在显著差异(P<0.05)。【结论】朝鲜鹌鹑MHC classⅠ基因第2、3外显子具有丰富的多态性,并且与新城疫病毒抗性相关。【Objective】 The study analyzed the allelic polymorphism of exons 1,2 and 3 of MHC class Ⅰ gene in Korean quail and the association with the Newcastle disease virus (NDV) resistance.【Method】 Based on conventional feeding method, fourteen-day-old Korean quails were challenged with La Sota attenuated strain of ND once a week by injection method.Three weeks after continuous challenge,blood samples were obtained from the heart.The β-micro method was used to determine the levels of NDV in serum.Amplification of MHC class Ⅰ gene in Korean quail by PCR technology,DNA Star and SPSS were used to analyze the allelic polymorphism of MHC class Ⅰ gene exons 2 and 3 and the NDV antibody differences among genotypes.【Result】 The ND antibody valence in Korean quail was 2 6-2 10 .PCR amplification obtained 1 290 bp DNA sequence and the coding region sequence was 780 bp,encoding 259 amino acids.DNA sequences of exons 1,2 and 3 of 76,272 and 274 bp were obtained by comparing with the known Japanese quail RNA (AB005528).No mutation was detected on ex on 1,while 11 moderate polymorphic mutations loci were found in exons 2 and 3 (4 mutations loci on exon 2:257 bp (A/C),267 bp (A/T),366 bp (A/G),and 500 bp (T/G);7 mutations loci on exon 3:795 bp (T/C), 798 bp (T/C),810 bp (T/G),830 bp (A/G),873 bp (A/T),927 bp (A/G),and 932 bp (C/G).The 267 bp (A/T),830 bp (A/G), 873 bp (A/T),and 932 bp (C/G) mutations led to amino acid changes.There were significant differences in ND antibody levels between the 500 bp (T/G),873 bp (A/T) and 932 bp (C/G) mutation loci ( P <0.05).【Conclusion】 The exons 2 and 3 of Korean quail MHC class Ⅰ gene had abundant polymorphism and close relationship with NDV resistance.

关 键 词：朝鲜鹌鹑 MHCclassⅠ基因 新城疫病毒抗性 

分 类 号：S837.2[农业科学—畜牧学]