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从四个部分纯化的输卵管信使核糖核酸合成全长互补脱氧核糖核酸。

Synthesis of full length cDNAs from four partially purified oviduct mRNAs.

作者信息

Buell G N, Wickens M P, Payvar F, Schimke R T

出版信息

J Biol Chem. 1978 Apr 10;253(7):2471-82.

PMID:632280
Abstract

Total poly(A)-containing RNA prepared from hen oviduct and centrifuged on an isokinetic sucrose gradient displays four peaks of optical absorbance. These have been identified by translation in vitro as lysozyme, ovomucoid, ovalbumin, and conalbumin mRNAs. Isolation and recentrifugation of the peaks results in partial purification of each mRNA. Molecular weights have been determined for the mRNAs on agarose gels containing 20 mM methylmercury hydroxide. Each mRNA possesses a number of apparently untranslated nucleotides ranging from approximately 900 bases for ovalbumin and conalbumin mRNAs to 200 bases for ovomucoid and lysozyme mRNAs. The mRNAs have been copied with avian myeloblastosis virus reverse transcriptase. Each mRNA with the exception of conalbumin gives rise to a high proportion of full length cDNA. Several parameters previously reported to influence the size distribution of cDNA had no effect on the length of cDNA made from any mRNA fraction. The proportion of full length copy does depend on the reverse transcriptase lot.

摘要

从母鸡输卵管制备的总含聚腺苷酸(poly(A))RNA,经等速蔗糖梯度离心后呈现出四个吸光度峰值。通过体外翻译已将它们鉴定为溶菌酶、卵类粘蛋白、卵清蛋白和伴清蛋白的信使核糖核酸(mRNA)。对这些峰值进行分离并再次离心可实现每种mRNA的部分纯化。已在含有20 mM氢氧化甲基汞的琼脂糖凝胶上测定了这些mRNA的分子量。每种mRNA都有许多明显未翻译的核苷酸,卵清蛋白和伴清蛋白mRNA约有900个碱基,卵类粘蛋白和溶菌酶mRNA则有200个碱基。这些mRNA已用禽成髓细胞瘤病毒逆转录酶进行了复制。除伴清蛋白外,每种mRNA都产生了高比例的全长互补脱氧核糖核酸(cDNA)。先前报道的几个影响cDNA大小分布的参数对任何mRNA组分产生的cDNA长度均无影响。全长拷贝的比例确实取决于逆转录酶批次。

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