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Cloning and application of the genes regulating fecundity in mammals

哺乳动物繁殖力调节基因的克隆及应用

基本信息

批准号：
07456121
负责人：
TOJO Hideaki
金额：
$ 4.74万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07456121/
关键词：
c-kit SCF shiba goat caprine SRY HMG RT-PCR cloning

项目摘要

(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.

(1)克隆了具有DBW表型的柴坝山羊c-kit基因的cDNA并进行了序列测定。一个单一的氨基酸（丙氨酸残基）插入在KI区域的毛色无关的山羊。Robson的二级结构预测预测丙氨酸插入可能将DKR转角转化为α-螺旋。（二）、从15周龄雌性柴山山羊小脑mRNA构建的cDNA文库中筛选出SCF cDNA，并利用RT-PCR方法从小脑mRNA中获得SCF cDNA，其中一种同工型gSCF 825与其他物种中的KL-1同工型一致。该基因编码区为822 bp，编码274个氨基酸残基的多肽。两种同种型gSCF 825和gSCF 741在胎儿和成人皮肤中表达。山羊c-kit基因KI区的丙氨酸插入对山羊和绵羊具有高度的特异性，分离到5种SCF cDNA亚型，其中包括2种新的亚型，表达gSCF 825和gSCF 741。(3)测定了4.9kb山羊SRY DNA及其从启动子区到3'非翻译区的碱基序列。山羊SRY基因启动子区含有多种具有潜在生物学意义的基序，包括TATA盒、CAAT盒、茎环、直接应答以及HMG盒家族蛋白和SP-1的结合基序。羊、人、牛和小鼠的HMG结构域的水热图显示出几乎相同的模式。4个物种SRY蛋白全长的亲水性图清楚地表明，小鼠Sry蛋白与其他物种的蛋白完全不同。