Gordon J I, Deeley R G, Burns A T, Paterson B M, Christmann J L, Goldberger R F
J Biol Chem. 1977 Nov 25;252(22):8320-7.
Administration of 17beta-estradiol to roosters induced the synthesis of vitellogenin in the liver. The mRNA that specifies this protein has been purified from the livers of estrogen-treated roosters and has been shown to have a molecular weight of 2.3 X 10(6) (Deeley, R.G., Gordon, J.I., Burns, A.T.H., Mullinix, K.P., Bina-Stein, M., and Goldberger R.F. (1977) J. Biol. Chem. 252, 8310-8319). In order to rigorously establish the identity of the polypeptide specified by this mRNA, we used a staphylococcal nuclease-treated, mRNA-dependent wheat germ cell-free translation system capable of synthesizing polypeptides as large as vitellogenin (monomer Mr = 240,000). Vitellogenin mRNA directs the in vitro synthesis of a polypeptide with the following features: (a) it co-migrates with authentic vitellogenin in SDS-polyacrylamide gels; (b) it is highly enriched for serine but is not phosphorylated; (c) it is immunoprecipitated by purified, monospecific, anti-vitellogenin antibody; and (d) it has an unusual cyanogen bromide cleavage pattern characteristic of vitellogenin. The most striking characteristic of the cyanogen bromide cleavage products is an extremely large polypeptide (Mr = 90,000) that contains two phosvitins. The kinetics of incorporation of serine and methionine into vitellogenin synthesized in the wheat germ cell-free translation system indicates that the phosvitins are located near the COOH-terminal portion of the molecule.
给公鸡注射17β-雌二醇可诱导肝脏中卵黄蛋白原的合成。编码这种蛋白质的mRNA已从经雌激素处理的公鸡肝脏中纯化出来，其分子量为2.3×10⁶（迪利，R.G.，戈登，J.I.，伯恩斯，A.T.H.，米利尼克斯，K.P.，比纳-斯坦因，M.，和戈德伯格，R.F.（1977年）《生物化学杂志》252，8310 - 8319）。为了严格确定由这种mRNA编码的多肽的身份，我们使用了一种经葡萄球菌核酸酶处理的、依赖mRNA的无细胞小麦胚芽翻译系统，该系统能够合成与卵黄蛋白原一样大的多肽（单体分子量 = 240,000）。卵黄蛋白原mRNA指导体外合成具有以下特征的多肽：（a）在SDS - 聚丙烯酰胺凝胶中它与天然卵黄蛋白原迁移率相同；（b）它富含丝氨酸但未磷酸化；（c）它能被纯化的、单特异性的抗卵黄蛋白原抗体免疫沉淀；（d）它具有卵黄蛋白原特有的不寻常的溴化氰裂解模式。溴化氰裂解产物最显著的特征是一个极大的多肽（分子量 = 90,000），它包含两个卵黄高磷蛋白。丝氨酸和甲硫氨酸掺入小麦胚芽无细胞翻译系统中合成的卵黄蛋白原的动力学表明，卵黄高磷蛋白位于分子的COOH末端附近。
