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诺维科夫肝癌、正常肝脏和再生肝脏的信使核糖核酸在互补脱氧核糖核酸-纤维素亲和基质上的吸附作用

Adsorption of messenger RNA of Novikoff hepatoma, normal liver, and regenerating liver on complementary DNA-cellulose affinity matrices.

作者信息

Hirsch F W, Nall K N, Hayes L C, Raju K S, Spohn W H, Busch H

出版信息

Cancer Res. 1977 Oct;37(10):3694-700.

PMID:71200
Abstract

Complementary DNA (cDNA)-oligodeoxythmidylate-celluloses were prepared from cDNA copies of polysomal messenger RNA (mRNA) of Novikoff hepatoma, normal rat liver, and regenerating rat liver. cDNA synthesis with reverse transcriptase was approximately 46% with respect to input mRNA with oligodeoxythymidylate-cellulose primer. The cDNA's of normal liver, regenerating liver, and Novikoff hepatomas were used as affinity matrices for hybridization of different mRNA species. Under the conditions used, degradation of mRNA was not detected. After normalization for homologous hybridization efficiency, 53 and 65% of the Novikoff hepatoma mRNA bound to normal liver and regenerating liver cDNA's. Under these conditions an average of 82% of mRNA of normal liver bound to regenerating liver cDNA, and 92% of regenerating liver mRNA bound to normal liver cDNA. The bound and unbound mRNA's were analyzed by translation in the wheat germ system; 2-D gel analysis of the proteins synthesized in the wheat germ system indicated that the cDNA affinity columns selectively adsorbed some mRNA species.

摘要

互补脱氧核糖核酸(cDNA)-寡聚脱氧胸苷酸-纤维素是由诺维科夫肝癌、正常大鼠肝脏和再生大鼠肝脏的多聚核糖体信使核糖核酸(mRNA)的cDNA拷贝制备而成。使用寡聚脱氧胸苷酸-纤维素引物,通过逆转录酶进行的cDNA合成相对于输入的mRNA约为46%。正常肝脏、再生肝脏和诺维科夫肝癌的cDNA用作不同mRNA种类杂交的亲和基质。在所使用的条件下,未检测到mRNA的降解。在对同源杂交效率进行标准化后,53%和65%的诺维科夫肝癌mRNA与正常肝脏和再生肝脏的cDNA结合。在这些条件下,平均82%的正常肝脏mRNA与再生肝脏cDNA结合,92%的再生肝脏mRNA与正常肝脏cDNA结合。通过在小麦胚芽系统中进行翻译来分析结合和未结合的mRNA;对在小麦胚芽系统中合成的蛋白质进行二维凝胶分析表明,cDNA亲和柱选择性地吸附了一些mRNA种类。

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