不同品种猪PBD-124基因多态性及差异表达研究  

Study on Polymorphism and Differential Expression of PBD-124 Gene in Different Breeds of Pigs

作　　者：高倍瑶 刘艳光 贾琪 柳俭强 罗新惠 张立春[2] GAO Beiyao;LIU Yanguang;JIA Qi;LIU Jianqiang;LUO Xinhui;ZHANG Lichun(Institute of Animal Science and Technology,Jilin Agricultural University,Changchun 130000,China;Institute of Animal Biotechnology,Jilin Academy of Agricultural Sciences,Gongzhuling 136100,China;College of Agriculture,Yanbian University,Yanji 130021,China)

机构地区：[1]吉林农业大学动物科技学院,长春130000 [2]吉林省农业科学院动物生物技术研究所,公主岭136100 [3]延边大学农学院,延吉130021

出　　处：《中国畜牧兽医》2022年第6期2208-2215,共8页China Animal Husbandry & Veterinary Medicine

基　　金：吉林省自然科学基金“民猪抗病标志因子筛选及验证”(20200201130JC)。

摘　　要：【目的】克隆获得猪β-防御素-124(porcine beta-defensin-124,PBD-124)基因CDS区并探究其多态性,分析PBD-124基因在不同品种猪及同种猪不同组织内的表达情况。【方法】采用RT-PCR方法扩增并克隆猪PBD-124基因CDS区,利用PCR-RFLP酶切法对大白猪、民猪和野杂猪PBD-124基因的BlnⅠ酶切位点进行多态性检测,利用实时荧光定量PCR方法检测该基因在不同品种猪肝脏、脾脏和血液内的表达差异。【结果】试验成功克隆出猪PBD-124基因CDS区,长423 bp,共编码140个氨基酸,测序结果发现其存在c.257 G>A和c.263 T>G 2个突变位点,因2个突变位点间隔过近,可能存在连锁,仅对第1个突变位点进行酶切,大白猪中检测到GG、GA、AA 3种基因型,而民猪和野杂猪中仅检测到GA和AA 2种基因型,3个群体中AA基因型均为优势基因型,大白猪、民猪和野杂猪的AA基因型频率分别为0.5261、0.9412和0.6452,且3个群体均处于Hardy-Weinberg平衡(P>0.05)。3个群体的多态性均不高,大白猪处于中度多态(0.25<PIC<0.5),民猪和野杂猪则处于低度多态(PIC<0.25)。实时荧光定量PCR结果显示,PBD-124基因在大白猪、民猪、野杂猪的脾脏、肝脏及血液中均有表达,且存在表达差异。【结论】猪PBD-124基因CDS区长423 bp,共编码140个氨基酸,与参考序列比对发现2个突变位点,均未引起氨基酸突变;3个群体多态性均不高;PBD-124基因在不同品种猪及同种猪不同组织间表达均存在差异。【Objective】The CDS region of porcine beta-defensin-124(PBD-124)gene was cloned and its polymorphism was explored.The expression of PBD-124 gene in different breeds of pigs and different tissues of the same kind of pigs was analyzed.【Method】The CDS region of PBD-124 gene was amplified by RT-PCR and cloned.PCR-RFLP was used to detect the polymorphism of PBD-124 gene(BlnⅠ)in Large White,Min and wild hybrid pigs.The expression difference of PBD-124 gene in liver,spleen and blood of different breeds of pigs was detected by Real-time quantitative PCR.【Result】The CDS region of PBD-124 gene was successfully cloned,with a length of 423 bp and a total of 140 amino acids.Sequencing results showed that there were two mutation sites of c.257 G>A and c.263 T>G.Because there was linkage between the two mutation sites,only the first mutation site was digested.Three genotypes of GG,GA and AA were detected in Large White pigs,while only two genotypes of GA and AA were detected in Min and wild hybrid pigs.AA genotype was the dominant genotype in three pig populations.The genotype frequency of Large White,Min and wild hybrid pigs were 0.5261,0.9412 and 0.6452,respectively,and three pig populations were in Hardy-Weinberg equilibrium(P>0.05).The polymorphism of three pig populations was not high.Large White pigs were in moderate polypeptide(0.25<PIC<0.5),while Min and wild hybrid pigs were in low polypeptide(PIC<0.25).Real-time quantitative PCR results showed that PBD-124 gene was expressed in spleen,liver and blood of Large White,Min and wild hybrid pigs,and there were expression differences.【Conclusion】The length of PBD-124 gene CDS was 423 bp,and encoded 140 amino acids.Compared with the reference sequence,two SNP mutation sites were found,which did not cause amino acid mutation.The polymorphism of three pig populations was not high.The expression of PBD-124 gene was different in different breeds of pigs and different tissues of the same kind of pigs.

关 键 词：大白猪 民猪 野杂猪 PBD-124基因 PCR-RFLP 组织表达 

分 类 号：S813.3[农业科学—畜牧学]