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来自2型腺病毒E3区域的三种剪接mRNA的结构

Structure of three spliced mRNAs from region E3 of adenovirus type 2.

作者信息

Stålhandske P, Persson H, Perricaudet M, Philipson L, Pettersson U

出版信息

Gene. 1983 May-Jun;22(2-3):157-65. doi: 10.1016/0378-1119(83)90099-9.

Abstract

A cDNA library representing early adenovirus type 2 (Ad2) mRNA was constructed. The cDNA copies were inserted into the PstI cleavage site of the pBR322 plasmid, and clones containing sequences from region E3 of the Ad2 genome were identified by colony hybridization. Selected clones were characterized by restriction enzyme cleavage, hybridization, and partial DNA sequence analysis. The precise structure of three spliced mRNAs was established by comparing the results with the DNA sequence of region E3 from Ad2 (Herissé et al., Nucl. Acids Res. 8 (1980) 2173--2191; Herissé and Galibert, Nucl. Acids Res. 9 (1981) 1229--1249). One of the characterized mRNA species encodes the E3/19K glycoprotein, whereas the other two most likely encode the E3/14K protein. The results demonstrate, moreover, that certain splice points which are used to generate the major E3 mRNAs are also used to splice the supplementary leader segments to the fibre mRNA at late times after infection. Two separate poly(A)-addition sites were identified in region E3 by analysis of the cDNA clones; one is preceded by the hexanucleotide sequence AAUAAA, whereas the other is preceded by an altered hexanucleotide, having the sequence AUUAAA.

摘要

构建了一个代表2型腺病毒(Ad2)早期mRNA的cDNA文库。将cDNA拷贝插入pBR322质粒的PstI切割位点,并通过菌落杂交鉴定含有Ad2基因组E3区序列的克隆。通过限制性内切酶切割、杂交和部分DNA序列分析对选定的克隆进行表征。通过将结果与Ad2 E3区的DNA序列进行比较,确定了三种剪接mRNA的精确结构(Herissé等人,《核酸研究》8(1980)2173 - 2191;Herissé和Galibert,《核酸研究》9(1981)1229 - 1249)。其中一种表征的mRNA物种编码E3/19K糖蛋白,而另外两种最有可能编码E3/14K蛋白。此外,结果表明,用于产生主要E3 mRNA的某些剪接位点在感染后期也用于将补充前导序列剪接到纤维mRNA上。通过对cDNA克隆的分析,在E3区鉴定出两个独立的聚腺苷酸化位点;一个位点之前是六核苷酸序列AAUAAA,而另一个位点之前是一个改变的六核苷酸,序列为AUUAAA。

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