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包含成年鸡和鸭珠蛋白信使序列的cDNA重组体的限制酶切图谱分析:一项比较研究。

Restriction mapping of cDNA recombinants including the adult chicken and duck globin messenger sequences: a comparative study.

作者信息

Reynaud C A, Ben Tahar S, Krust A, Amaral de Lima Franco M P, Goldenberg S, Gannon F, Scherrer K

出版信息

Gene. 1980 Nov;11(3-4):259-69. doi: 10.1016/0378-1119(80)90066-9.

Abstract

A comparison of the organization of six avian adult globin messenger sequences is based on previously reported recombinant duck adult globin cDNA plasmids (Therwath et al., 1980) and the actual construction and characterization of pBR322 recombinant plasmids including the beta and the normal alpha A and alpha D chicken adult globin mRNA sequences. Identification of the cloned DNA was performed using hybridization-selection under conditions permitting complete purification in one step of the three globin mRNAs, and translation of the corresponding mRNA. Orientation of the globin insert in the vector was determined, taking into account the computer prediction of the restriction sites based on the known amino acid sequences of the three globin chains (Roizès and Pelaquier, 1980) and those actually observed, and by identification of restriction fragments using 3'-specific probes. Identification, orientation and restriction mapping of these cloned DNAs reveals extensive homologies in organisation of beta sequences between duck and chicken, as well as among the alpha sequences in every two possible combinations.

摘要

基于先前报道的重组鸭成体珠蛋白cDNA质粒(Therwath等人,1980年)以及包含β链、正常αA和αD鸡成体珠蛋白mRNA序列的pBR322重组质粒的实际构建和表征,对六种鸟类成体珠蛋白信使序列的组织进行了比较。使用杂交选择法进行克隆DNA的鉴定,该方法在允许一步完全纯化三种珠蛋白mRNA并翻译相应mRNA的条件下进行。考虑到基于三种珠蛋白链已知氨基酸序列(Roizès和Pelaquier,1980年)以及实际观察到的限制酶切位点的计算机预测,并通过使用3'特异性探针鉴定限制酶切片段,确定了载体中珠蛋白插入片段的方向。这些克隆DNA的鉴定、方向确定和限制酶切图谱分析揭示了鸭和鸡之间β序列组织以及每两种可能组合的α序列之间的广泛同源性。

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