普定高脚鸡甲状旁腺激素相关蛋白基因PTHrP多态性及组织差异表达分析  

Polymorphism and Tissue Expression Pattern of Parathyroid Hormonerelated Peptide Gene(PTHrP) in Gaojiao Chicken(Gallus)

作　　者：杨红[1,2] 孙鹃[3] 张勇[1,2] 裘贵华 李俊[1,2] 张雄[1,2] 彭松[1,2] 

机构地区：[1]贵州大学高原山地动物遗传育种与繁殖省部共建教育部重点实验室,贵阳550025 [2]贵州大学动物科学学院贵州省动物遗传育种与繁殖重点实验室,贵阳550025 [3]贵州省畜禽遗传资源管理站,贵阳550001

出　　处：《农业生物技术学报》2015年第8期1084-1092,共9页Journal of Agricultural Biotechnology

基　　金：贵州省农业动植物育种专项项目<高脚鸡配套系选育及推广应用>(项目编号:黔农育专字[2012]10号);浙江大学2012年基本科研业务费专项资金课题--<贵州畜禽遗传资源库平台建设>(项目编号:2012XZZX003-1)

摘　　要：为揭示普定高脚鸡甲状旁腺激素相关蛋白基因(parathyroid hormone-related peptide gene,PTHr P)的遗传多样性及其在各组织中的差异表达,本实验以贵州省特有的高脚鸡为实验材料,矮脚鸡和百宜黑鸡作对比,利用DNA池、单链构象多态性(single strand conformation polymorphim,SSCP)和DNA测序技术对高脚鸡PTHr P进行了多态位点检测;采用实时荧光定量PCR技术对PTHr P在高脚鸡、矮脚鸡和百宜黑鸡的心脏、肝脏、胸肌、腿肌中的差异表达进行了分析。结果表明,高脚鸡PTHr P在第3外显子209 bp处发生了C→T突变,为同义突变,SSCP检测出3种基因型CC、CT和TT,相关分析显示C209T多态位点对高脚鸡胫长具有显著影响,且CT基因型个体胫长均显著高于CC和TT基因型个体(P<0.05),经χ2适合性检验表明高脚鸡群体呈现中度多态(PIC=0.350),并处于Hardy-weinberg平衡状态(P>0.05);RT-PCR检测显示PTHr P在高脚鸡腿肌中表达量最高,且显著高于其他组织(P<0.05);在与矮脚鸡、百宜黑鸡比较中,高脚鸡各组织中的表达量均高于百宜黑鸡相应组织表达量(P<0.05),但均低于矮脚鸡相应组织表达量(P<0.05)。研究结果提示,PTHr P基因多态性与高脚鸡胫长显著相关,可尝试用CT基因型为筛选高脚鸡胫长的分子标记辅助选择,且PTHr P的表达量在一定程度上对鸡胫骨长度具有影响作用。In order to reveal the genetic diversity and tissue expression pattem of Parathyroid hormone- related peptide gene （PTHrP） in Gaojiao chickens （Callus）, Gaojiao chickens in Guizhou province were used in the present study, Aijiao chickens and Baiyi black chickens as controls. Its polymorphic loci were detected with DNA pool, Single Strand Conformation Polymorphism （SSCP） and DNA sequencing techniques, and its differential expression levels in heart, liver, breast muscl and leg muscle were analyzed by using real-time quantitative PCR technology. The results showed that the C→T point mutation was detected in 209bp of exon 3 of PTHrP gene of Gaojiao chickens, which was a silent mutation, hree genotypes （CC, CT and TT） werefound.The relationship between PTHrP gene and shank length in Gaojiao chickens was analyzed. The C209T polymorphism wad significantly associate with shank length of Gaojiao chicken, and the individuals with CT genotype had significant shank length than CC and TT （P〈0.05）. Thefitness test indicated that Gaojiao chickens at the mutation locus were Hardy-weinberg equilibrium（P〉0.05）, and itpresented moderate polymorphism （PIC=0.350）. The RQ-PCR detection showed that the expression level of PTHrP gene was the highest in leg muscle of Gaojiao chickens, and significant higher than other tissues （P〈0.05）. In addition, compare with Aijiao chicken and Baiyi black chicken,the expression levels of various tissues of Gaojiao chickens were higher than that of Baiyi black chickens （P〈0.05）, but lower than Aijiao chickens （P^0.05）. The results indicate that polymorphisms of PTHrP gene are significantly correlated with shank length traits of Gaojiao chickens, and individuals with CT genotype can be used as molecular genetic marker to select shank length of Gaojiao chicken for reproduction traits, and the expression levels of PTHrP has an influence on chickens＇ shank length.

关 键 词：高脚鸡 PTHr P基因 PCR-SSCP 荧光定量PCR 多态性 差异表达 

分 类 号：S634.1[农业科学—蔬菜学]