卤虫隐生胚囊中含聚腺苷酸信使核糖核蛋白的蛋白质成分的表征
Characterization of protein components of poly(A)-containing messenger ribonucleoproteins from cryptobiotic gastrulae of Artemia salina.
作者信息
Slegers H, De Herdt E, Kondo M
出版信息
Mol Biol Rep. 1979 May 31;5(1-2):65-9. doi: 10.1007/BF00777490.
Conditions for the isolation of intact poly(A)+mRNP from cryptobiotic gastruale of A. salina are described. In the presence of Mg2+ ions nucleolytic cleavage occurs in vitro in the vicinity of the 3'-poly(A) segment of mRNP during the isolation procedure. The resulting two parts of poly(A)+mRNP complex are separated by thermal elution from oligo(dT)-cellulsoe affinity column. Analysis by SDS-gel electrophoresis of protein components associated with intact poly(A)+mRNP has revealed the existence of 20--30 S RNP complex containing five major proteins with Mr 68,000, 53,000, 50,000, 45,000 and 38,000, respectively, but completely lacking the poly(A)-specific Mr 76,000 protein.
本文描述了从盐生丰年虫隐生胚中分离完整的聚腺苷酸（poly(A)）+信使核糖核蛋白（mRNP）的条件。在分离过程中，在镁离子（Mg2+）存在的情况下，mRNP的3'-聚腺苷酸（poly(A)）片段附近会发生体外核酸酶切割。通过从寡聚（dT）-纤维素亲和柱上进行热洗脱，将所得的聚腺苷酸（poly(A)）+mRNP复合物的两部分分离。对与完整聚腺苷酸（poly(A)）+mRNP相关的蛋白质成分进行十二烷基硫酸钠-凝胶电泳（SDS-凝胶电泳）分析，结果显示存在一种20-30S的核糖核蛋白（RNP）复合物，该复合物包含五种主要蛋白质，其分子量（Mr）分别为68,000、53,000、50,000、45,000和38,000，但完全缺乏聚腺苷酸（poly(A)）特异性的分子量为76,000的蛋白质。
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