【作者】 滑国华；

【导师】 陈世林； 杨利国；

【作者基本信息】 华中农业大学 ， 动物遗传育种与繁殖， 2006， 硕士

【摘要】 在现代动物育种实践中，应用分子标记辅助选择（Marker-assisted selection，MAS）对低遗传力性状、伴性性状或表达较晚的性状进行选育具有非常重要的意义。利用这种方法进行山羊繁殖性状改良和选择的前提，是必须找到尽可能多的分子标记。 本研究选取骨形态发生蛋白受体IB基因（Bone morphogenetic protein receptor IB，BMPR-IB）、骨形态发生蛋白15基因（Bone morphogenetic protein 15，BMP15）、抑制素a亚基基因（Inhibin-a subunit gene，INHA）、褪黑激素1型受体a亚型基因(Mel_1amelatonin receptor，MTNR1A)和促卵泡素β亚基基因（Follicle stimulating hormone β，FSHβ）为候选基因，以及与FecB（Booroola Fecundity）基因连锁的3个微卫星位点BMS2508、BM143和OARHH35，分析它们在波尔山羊（209只）、海门山羊（54只）、努比山羊（37只）和马头山羊（23只）4个山羊品种共323个样本中的多态性，并对不同基因型与产羔性状的关系进行了探索。主要研究结果如下： 1．利用Forced PCR-RFLP技术证实：在所检测的波尔山羊、海门山羊、马头山羊和努比山羊4个山羊群体中没有发现FecB基因突变和BMP15基因的FecX^I、FecX^H、FecX^G、FecX^B四个点突变。 2．在4个山羊品种抑制素a亚基基因（INHA）中发现G¹²⁸-A¹²⁸（Accession number：L28815）突变，该突变引起Hae Ⅱ酶切位点改变；利用PCR-RFLP技术进行基因型判定，发现波尔山羊和努比山羊基因型频率差异显著（P＜0.05），努比山羊与海门山羊基因型频率差异极显著（P＜0.01）；G等位基因可能与山羊高产性状呈正相关。 3．在4个山羊品种褪黑激素1型受体a亚型基因（MTNR1A）中共发现7个点突变，分别是AY524665序列T²⁴⁶-C²⁴⁶突变（引起Xba Ⅰ酶切位点改变）和T²⁷⁸-C²⁷⁸突变，AF419334序列T¹⁵⁵-C¹⁵⁵突变、A²³⁹-G²³⁹突变、A²⁷⁷-G²⁷⁷突变、G³⁵³-A³⁵³突变（引起Bst4CI酶切位点改变）和T⁴²⁵-C⁴²⁵突变，对其中4个点突变进行了基因型判定，其中三个位点(AF419334序列T¹⁵⁵-C¹⁵⁵突变、A²³⁹-G²³⁹突变、A²⁷⁷-G²⁷⁷突变)用AS-PCR方法进行基因型判定，AY524665序列T²⁴⁶-C²⁴⁶突变用Xba Ⅰ-RFLP进行基因型判定。 4．4个山羊品种褪黑激素1型受体a亚型基因（MTNR1A，Accession number：AF419334）T¹⁵⁵-C¹⁵⁵突变用AS-PCR技术判定基因型，并进行性状关联分析，结果发现：波尔山羊和海门山羊基因型频率差异极显著（P＜0.01），其它品种间差异不显著；A₁等位基因可能与海门山羊和波尔山羊产羔数呈正相关。 5．4个山羊品种褪黑激素1型受体a亚型基因（MTNR1A，Accession number：AF419334）A²³⁹-G²³⁹突变用AS-PCR技术判定基因，并进行相关分析，结果发现：更多还原

【Abstract】 In the modern animal breeding practice, it’s very important to improve breeding efficiency for sex linked, low heritability or late-expressive traits by Marker-assisted selection （MAS）. Before using this method, we must find suitable molecular markers as many as possible.Polymorphisms of BMPR-IB （Bone morphogenetic protein receptor IB）, BMP15 （Bone morphogenetic protein 15）, INHA （Inhibin-a subunit gene）, MTNR1A (Mel_1a melatonin receptor) and FSHβ （Follicle stimulating hormone 6） were studied as candidate genes in Boar goat （209）, Haimen goat （54）, Nubi goat （37） and Matou goat （23） in addition with 3 microsatellite loci BMS2508, BM143 and OARHH35 linked with FecB gene and their effects on litter size were analyzed. The main results were showing as follows:1. Mutations in FecB and FecX^I, FecX^H, FecX^G and FecX^B in BMP15 were not found in the four investigating goat breeds such as Boar, Haimen, Nubi goat and Matou goat by Forced PCR-RFLP.2. Polymorphism of INHA was studied in Boar goat, Haimen goat, Nubi goat and Matou goat. It was the first time to discover a new mutation G¹²⁸ -A¹²⁸ （Accession number: L28815） in the goat. A Hae II polymorphism was detected at this site by PCR-RFLP analyzing. There was significant differences on the genotype frequencies between Boar and Nubi goat （P<0.05）, between Nubi and Haimen goat at P<0.01. The allele G was positively correlated with litter size possibly.3. Seven point mutations were discovered in MTNR1A in these 4 goat breeds, that were T-C at 246, 278 （Accession number: AY524665） and 155, 425 （Accession number: AF419334）, A-G at 239, 277 and 353 （Accession number: AF419334）. Polymorphisms was detected at 155, 239 and 277 （Accession number: AF419334） by AS-PCR. Xba I polymorphism was detected at 246 （Accession number: AY524665） by PCR-RFLP.4. Polymorphism at 155 was detected （Accession number: AF419334） by AS-PCR and its effects on litter size were studied. Results showed that genotype frequencies更多还原