鸡BF基因第2、3外显子序列及其多态性分析  

Sequence and polymorphism of exons 2 and 3 of BF gene in chicken

作　　者：王威 勾越 金元昌[1] 张舟 任朝云 陈修月 宋国娇 

机构地区：[1]湖南科技大学生命科学学院,湖南湘潭411201

出　　处：《中国生物制品学杂志》2017年第12期1269-1273,共5页Chinese Journal of Biologicals

基　　金：湖南省教育厅重点项目(15A067);生物工程专业省级校企合作人才培养示范基地(G145813)

摘　　要：目的分析鸡BF基因第2、3外显子序列及其多态性。方法从6羽霞烟鸡外周血淋巴白细胞(peripheral blood leukocytes,PBLs)中提取细胞DNA,以其为模板,经PCR法扩增BF基因的第2外显子、第2内含子和第3外显子,克隆至pGM-T载体上,构建重组质粒pGM-T-BF,并进行PCR、酶切和测序鉴定。利用生物信息学DNAStar软件将霞烟鸡与红原鸡第2、3外显子核苷酸及氨基酸序列进行多序列比对。结果经PCR、酶切及测序证明重组质粒pGM-T-BF构建正确。2种鸡BF基因序列的同源性可达88.6%~100%,第2、3外显子存在多态性变异位点91个,占总长度的17.04%,其中简约性信息位点66个,单变异位点25个;在编码相应的178个氨基酸残基中,多态变异位点49个,占总氨基酸残基数的27.53%,其中简约性信息位点37个,分单变异位点12个。结论鸡BF基因第2、3外显子呈高度多态性,且其多态性更多表现在氨基酸水平上。Objective To analyze the sequence and polymorphism of exons 2 and 3 of BF gene in chicken. Methods DNA was isolated from the peripheral blood leukocytes（PBLs） of six Xiayan chickens and used as a template for amplification of exon 2, intron 2 and exon 3 of BF gene by PCR. The PCR products were cloned into vector pGM-T, and the constructed recombinant plasmid pGM-T-BF was identified by PCR, restriction analysis and sequencing. Bioinformatics software DNAStar was used to analyze multiple alignment of nucleotide and amino acid sequences of exons 2 and 3 of Xiayan chickens with those of Red junglefowls. Results PCR, restriction analysis and sequencinmg proved that recombinant plasmid pGM-T-BF was constructed correctly. The homologies of BF genes of Xiayan chickens to those of Red junglefowls were 88. 6% ~ 100%. There were 91 diversity sites in exons 2 and 3, which accounted for 17. 04% of total length, including 66 parsim-info sites and 25 singleton sites. However, there were 49 diversity sites in 178 amino acid residues, which accounted for 27. 53%, including 37 parsim-fnfo sites and 12 singleton sites. Conclusion The exons 2 and 3 of chicken BF gene showed high diversity, epecially at amino acid level.

关 键 词：主要组织相容性复合体 外显子 多态性 核苷酸序列 氨基酸序列 霞烟鸡 

分 类 号：Q789[生物学—分子生物学]