Cloning and application of the genes regulating fecundity in mammals

(1)克隆并测序了具有DBW表型的柴山羊c-kit cDNA。在不同毛色的山羊中，KI区存在一个氨基酸（丙氨酸残基）插入。罗布森二级结构预测预测丙氨酸的插入可能使DKR转向a-螺旋。（2）. 通过筛选15周龄柴山羊母羊小脑mRNA cDNA文库克隆SCF cDNA，并采用RT-PCR方法从小脑mRNA中获得SCF cDNA。其中一种异构体gSCF825与其他物种的KL-1异构体相对应。它具有822bp的编码区，可编码长度为274个氨基酸残基的肽。gSCF825和gSCF741两种异构体在胎儿和成人皮肤中均有表达。绵羊c-kit KI区的丙氨酸插入对山羊和绵羊具有高度特异性，分离出5个SCF cDNA亚型，其中包括2个新的亚型，在柴巴山羊皮肤中表达了gSCF825和gSCF741两个亚型。(3)确定了4.9 kb的山羊SRY DNA及其启动子区至3′非翻译区碱基序列。山羊SRY的启动子区域包含多种具有潜在生物学意义的基序，包括TATA盒、CAAT盒、茎环、直接重复以及HMG-box家族蛋白和SP-1的结合基序。山羊、人、牛和小鼠的HMG结构域的水化图表现出几乎相同的模式。4种SRY蛋白全长度的亲水图清楚地表明，小鼠SRY蛋白与其他物种完全不同。

(1) The c-kit cDNA of Shiba goats with the DBW phenotype was cloned and sequenced. A single amino acid (alanine residue) insertion was presented in the KI region irrespective of coat color in goats. The Robson's prediction of the secondary structure predicted the alanine insertion might convert the DKR turn to a-helix. (2). SCF cDNAs were cloned by screening a cDNA library constructed from cerebellar mRNA of a 15-week-old female Shiba goat, RT-PCR methods were also used to obtain SCF cDNA from cerebellar mRNA.One of the isoforms, gSCF825, corresponds to KL-1 isoform in other species. It has 822bp codingregion and would encode peptide of 274 amino acid residues in length. Two isoforms, gSCF825 and gSCF741, are expressed in both fetal and adult skin. The alanine insertion in the KI region of caprine c-kit is highly specific to goats and sheep, fiveisoforms of SCF cDNA including2 novel isoforms were isolated, and in the skin of Shiba goats, two isoforms i.e., gSCF825 and gSCF741 were expressed. (3) The 4.9 kb caprine SRY DNA and its base sequence from the promotor region to the 3' non-translated region were determined. The promoter region of caprine SRY contained various motifs of potential biological significance including, TATA box, CAAT box, stem loop, direct reoeats and the binding motifs for HMG-box family proteins and SP-1. The hydrophathyplots of HMG domains of goat, human, cattle and mouse exhibited almost the identical patterns. The hydropathy plots of the entire length of SRY proteins of the 4 species demonstrated clearly that the murine Sry protein is completely different from those of other species.