摘要
从牛垂体中分离出总mRNA,经逆转录酶及大肠杆菌DNA聚合酶合成双链cDNA,经T4DNA聚合酶切成平末端后与Sma I酶切的pUC19连接,转化JM83,构建了脑垂体mRNA的cDNA文库。用标记的牛促卵泡激素基因的寡核苷酸片段进行杂交,从文库中筛选到几个阳性克隆,其中一个含有全长的牛促卵泡激素基因编码序列。利用PCR扩增技术从cDNA中扩增出了387bp的牛促卵泡激素基因,序列分析表明,10号克隆中的牛促卵泡激素基因含有起始密码ATG及终止密码TAA,所编码的氨基酸序列与天然牛促卵泡激寡素的氨基酸序列相同。
Total mRNA of Bovine Pituitary was isolated and purified. The eDNA library was then constructed. Bovine follicle stimulating hormone (FSH) gene was screened using synthetic oligonucleotide probe and Polymerase Chain Reaction (PCR). By using PCR technique, one 387bp DNA fragment was amplified from eDNA, which was then cloned into pUC19, M13mp18 and M13mp19 for DNA sequence analysis, results show that the DNA fragment is coding for a FSH of 109 amino acids and a signal peptide of 20 amino acids which sequence is the same as the published one.
出处
《西北农业学报》
CAS
CSCD
1992年第1期4-7,T001,共5页
Acta Agriculturae Boreali-occidentalis Sinica
关键词
促卵泡激素
CDNA克隆
序列分析
牛
Follicle stimulating hormome
cDNA clone
PCR
Sequence analysis
