Cloning and application of the genes regulating fecundity in mammals

(1)对具有DBW表型的柴巴山羊的c-kit基因进行了克隆和测序。无论山羊的毛色如何，KI区均存在单一氨基酸(丙氨酸残基)插入。Robson对二级结构的预测预测，丙氨酸插入可能会将DKR转向a-螺旋。(2)。通过筛选15周龄柴巴母羊小脑基因文库，克隆了SCF基因，并用RT-PCR方法从小脑中获得了SCF基因，其中一个亚型为gSCF825，与其他物种中的KL-1亚型相对应。该基因编码区822bp，编码274个氨基酸残基的多肽。两种异构体gSCF825和gSCF741在胎儿和成人皮肤中都有表达。山羊和绵羊在c-kit的K1区插入丙氨酸是高度特异的，共分离到包括2个新亚型在内的5种SCF亚型，在柴胡山羊皮肤中表达了两种亚型，即gSCF825和gSCF741。(3)测定了4.9kb的山羊SRY DNA及其从启动子区域到3‘非翻译区的碱基序列。山羊SRY基因启动子区域包含多种具有潜在生物学意义的基序，包括TATA box、CAAT box、茎环、直接反应序列以及与HMG-box家族蛋白和SP-1结合的基序。山羊、人、牛和小鼠的HMG结构域的水相图显示了几乎相同的模式。4个物种SRY蛋白全长的亲水性图谱表明，小鼠Sry蛋白与其他物种的SRY蛋白完全不同。

(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.