牛泌乳素mRNA的分离及鉴定  

Isolation and Characterization of Prolactin Messenger RNA from Bovine Anterior Pituitary Glands

作　　者：张同海[1] 宋诗铎[1] 李立津[1] 祁伟[1] 胡文芝[1] 王培福[1] 陈昆明[2] 方佩华[2] 

机构地区：[1]天津医学院第二附属医院,300210 [2]天津市内分泌研究所

出　　处：《Acta Genetica Sinica》1992年第5期410-415,共6页

基　　金：国家自然科学基金~~

摘　　要：本文报道了从牛脑垂体提取总RNA,经寡聚脱氧胸苷纤维素亲合层析分离获得牛脑垂体Poly(A)^+RNA。牛泌乳素mRNA经含羟甲基汞琼脂糖凝胶电泳分析,估计其长度约为1200个核苷酸。根据牛泌乳素的部分氨基酸序列推断并合成寡聚核苷酸探针,经Northern印迹杂交及放射自显影分析,证实了该mRNA中含有牛泌乳素mRNA的序列。在兔网织红细胞体外翻译体系中,牛泌乳素mRNA促进了~35S-甲硫氨酸参入,翻译合成的初级翻译产物能与兔抗羊PRL抗血清发生特异性免疫沉淀反应,SDS聚丙烯酰胺凝胶电泳及放射自显影分析结果表明,牛泌乳素前体的分子量约25000。The messenger RNA was extracted from bovine anterior pituitary glands and was purified by oligo(dT)-cellulose chromatography. The length of the mRNA was 1 200 nucleotides measured by agarose gel electrophoresis containing methylmercury hydroxide. The bovine prolaetin (PRL) mRNA was confirmed by Northern blot analysis of the mRNA with γ-22P labelled synthetic oli-gonucleodde probes based on the partial amino acid sequences of bovine PRL and could stimulate the incorporation of 25S-methionine into protein in the rabbit reticulocyte cell-free system. The translation product of bovine PRL mRNA was immunoprccitable by rabbit anti-ovine PRL an-riantibodies. The molecular weight of the translation product corresponding to the bovine PRL precursor was estimated to be approxiately 25 000 by SDS polyacrylamide gel electrophoresis and autoradiograph.

关 键 词：牛泌乳素 MRNA 分离 鉴定 

分 类 号：Q342[生物学—遗传学]