Noce T, Ando H, Ueda T, Kubokawa K, Higashinakagawa T, Ishii S
Department of Developmental Biology, Mitsubishi Kasei Institute of Life Sciences, Tokyo, Japan.
J Mol Endocrinol. 1989 Sep;3(2):129-37. doi: 10.1677/jme.0.0030129.
A cDNA expression library was constructed from poly(A)+ RNA of broiler chicken adenohypophyses using lambda gt11 as a vector. After screening with a rabbit antiserum against chicken LH, a cDNA clone (L12) containing a 436 bp insert was obtained. Using a subclone of L12 in pUC19 (pL12) as the hybridization probe, another cDNA clone (LF127) with a 533 bp insert was isolated. The LF127 contained the full-length cDNA encoding the putative chicken LH-beta subunit precursor molecule. Hybridization of the pL12 cDNA insert to adenohypophysial RNA showed that chicken and Japanese quail adenohypophyses contained RNA species of about 0.8 and 1.0 kb respectively. The amount of this RNA species was ten times higher in adult male quails kept under long days at room temperature than in those kept under short days at 7 degrees C. In situ hybridization experiments showed the exclusive distribution of the signal in the LH cells of the adenohypophysis. The similarity of the nucleotide sequence of the apoprotein-coding region of LH-beta cDNA of the chicken to that of mammals is lower than that among mammals. The deduced amino acid sequence of the chicken LH-beta subunit supports the hypothesis that the number of proline residues increases in the LH-beta subunit the closer phylogenetically the vertebrate is to mammals.
以λgt11为载体,从肉仔鸡腺垂体的聚腺苷酸加尾RNA构建了一个cDNA表达文库。用抗鸡促黄体激素的兔抗血清筛选后,获得了一个含有436 bp插入片段的cDNA克隆(L12)。以pUC19中的L12亚克隆(pL12)作为杂交探针,分离出另一个含有533 bp插入片段的cDNA克隆(LF127)。LF127包含编码假定的鸡促黄体激素β亚基前体分子的全长cDNA。pL12 cDNA插入片段与腺垂体RNA的杂交显示,鸡和日本鹌鹑的腺垂体分别含有约0.8 kb和1.0 kb的RNA种类。在室温下长日照饲养的成年雄性鹌鹑中,这种RNA种类的量比在7℃短日照饲养的鹌鹑中高10倍。原位杂交实验显示信号仅分布在腺垂体的促黄体激素细胞中。鸡促黄体激素β cDNA载脂蛋白编码区的核苷酸序列与哺乳动物的相似性低于哺乳动物之间的相似性。鸡促黄体激素β亚基的推导氨基酸序列支持这样的假设,即促黄体激素β亚基中脯氨酸残基的数量在系统发育上越接近哺乳动物的脊椎动物中增加得越多。