【作者】 白杰；

【导师】 李飞； 史洪才；

【作者基本信息】 石河子大学 ， 动物遗传育种与繁殖， 2007， 硕士

【摘要】 本研究以BMPR-IB基因,BMP15基因,GDF9基因和ESR基因为多胎候选主效基因,分别对策勒黑羊(167只),多浪羊(49只)进行了分析研究,结果表明,策勒黑羊和多浪羊均存在BMPR-IB基因的746单碱基A-G突变。策勒黑羊中三种基因型纯合子(BB)、杂合子(B+)、野生型(++)的频率分别是0.10(17)、0.35(58)和0.55(91),产羔数与基因型之间的分析结果表明,三种基因型之间的产羔数差异显著,其中++型和B+差异显著(p<0.05);++型和BB差异极显著(p<0.01)。策勒黑羊上没有检测到BMP15基因突变(FecXG、FecXB、FecXI、FecXH)和GDF9基因突变(FecGH),策勒黑羊ESR基因的第一外显子发生一个点突变(C-G突变),突变纯合子(EE)、杂合子(E+)和野生型(++)基因型频率分别是0.03(5 )、0.31(52)和0.66(110);对ESR基因型和策勒黑羊产羔数分析结果表明,策勒羊三种基因型之间产羔数差异显著,其中++型和EE差异显著(p<0.05)。通过BMPR-IB基因和ESR基因叠加效应分析表明,同时存在两种基因突变的个体的产羔数没有变化。从多浪羊460只样本中,检测到BMPR-IB的突变基因,++、B+频率分别为0.94和0.06,没有发现BB型个体,多浪羊(49只)没有发现BMP15基因和GDF9基因突变。通过上述研究,确定在新疆多浪羊和策勒黑羊中存在多胎主效基因BMPR-IB基因突变,策勒黑羊中还存在ESR基因突变,为今后在多浪羊和策勒黑羊中进行标记辅助选择奠定了分子生物学基础。根据点突变扩增的特点,结合国内外研究,建立了实验室大批量检测突变标记位点的Tetra-ARMS PCR技术,并对多浪羊460和策勒黑羊167个样本进行了检测,并与PCR-RFLP结果进行了对比,结果表明该技术有方便、快速、经济等优点,结果可靠,适合推广应用。更多还原

【Abstract】 BMPR-IB gene as a fecundity candidate gene was studied in Cele black sheep(167 blood samples) and Duolang sheep(509) by Tetra-primer ARMS-PCR technique. The results showed that there was a mutation (A746G) of BMPR-IB gene in Cele black sheep and Duolang sheep, between them, the BB、B+ and ++ frequence in Cele black sheep were 0.10、0.35 and 0.55 respectively. There was no BB genetype in detected Duolang sheep, and the B+、++ frequence were 0.06 and 0.94 respectively. Analyzing the association with litter size showed that, Cele black sheep with genotype BB had a most significantly difference litter size than those with genotype ++ (P<0.01); B+ had a significantly difference litter size than those with genotype ++ (P<0.05); BMPR-IB gene was a major gene influencing prolificacy in Cele black sheep; the Tetra-primer ARMS-PCR technique was simple,rapid and economical for detecting BMPR-IB mutation and other SNP locus.BMP15 and GDF9 as candidate gene of high fecundity were detected in Cele black sheep and Duolang sheep.No mutant genetypes were identified in FecG H lacus of GDF9 and FecXG、FecXB、FecXI、FecXH four locus of BMP15 .Our results suggested that BMP15 and GDF9 may not be the major gene for the high prolificity of Cele Black sheep and Duolang sheep.ESR gene as a fecundity candidate gene was studied in Cele black sheep(44 blood samples), Duolang sheep(176), Hu sheep (31) and Small Tail Han sheep (33blood samples) by PCR-RFLP and PCR-SSCP technique. The results showed that there was a mutation (C363G) of exon 1 of ESR gene in Cele black sheep, Hu sheep and Small Tail Han sheep . Among them, the EE、E+ and ++ frequence in Cele black sheep were 0.03、0.31 and 0.66 respectively; the EE、E+ and ++ frequence in Hu sheep were 0.03、0.19 and 0.77 respectively. And the EE、E+ and ++ frequence in Small Tail Han sheep were 0.03、0.42 and 0.55 respectively. Analyzing the association with litter size showed that, Cele black sheep with genotype EE had a most significantly difference litter size than those with genotype ++ (P<0.05); ESR gene was a major gene influencing prolificacy in Cele black sheep.更多还原