北京鸭卵对蛋白mRNA的提纯及某些性质的鉴定

 本文报道了从产卵期北京鸭输卵管中提取总RNA,经Olilo(dT)-纤维素柱层析,再经sepharose 4B柱层析步骤,得到了纯化的鸭卵清蛋白mRNA。我们建立了麦胚无细胞体系并探索了鸭卵清蛋白mRNA在此体系中翻译的最适条件。在此条件下测定了各纯化步骤的总mRNA翻译活性,并用免疫沉淀法测定其中卵清蛋白mRNA的活性。测定结果表明我们从mRNA中分离得到了纯鸭卵清蛋白mRNA。用变性的琼脂糖凝胶电泳对各纯化步骤的核酸样品进行组成分析,确定出鸭卵清蛋白mRNA的大小约为21S。

PURIFICATION AND PRELIMINARY CHARACTERIZATION OF OVALBUMIN MESSENGER RNA FROM PEKING DUCK

Preparation of purified ovalbumin mRNA was achieved by using combination of several techniques. Total RNA was extracted from the peking duck oviduct. Poly(A)containing mRNA was isolated from the total RNA extract by on oligo(dT) cellulose affinity at room temperature, and then was fractionated by gel filtration on a sepharose 4B column at 4℃ to obtain purified ovalbumin mRNA.For assaying the translation activety of duck ovalbumin mRNA, a wheat germ cell-free protein synthesezing system was established. The purity of the ovalbumin mRNA was assessed by determination of the relative translation activity of the total mRNA and ovalbumin mRNA in the wheat germ cell-free system. 100% of the total peptides synthesized were found to be specifically immunoprecipitable with an ovalbumin antiserum. The size of the duck ovalbumin mRNA characterized by denaturing agarose gel electrophoresis was 21s.