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一种高等植物核DNA序列的组织:大豆核糖体DNA的分子克隆与特性分析

The organization of a nuclear DNA sequence from a higher plant: molecular cloning and characterization of soybean ribosomal DNA.

作者信息

Varsanyi-Breiner A, Gusella J F, Keys C, Housman D E, Sullivan D, Brisson N, Verma D P

出版信息

Gene. 1979 Nov;7(3-4):317-34. doi: 10.1016/0378-1119(79)90051-9.

Abstract

The recombinant DNA vector, lambda Charon 4A, was used to construct a library of DNA sequences from the genomic DNA of soybean (Glycine max). To define the organization of ribosomal DNA (rDNA) in the soybean genome, clones containing sequences complementary to both 17S and 25S rRNA have been isolated from this library and used in conjunction with Southern blot hybridization. The rRNA genes are tandemly reiterated with a relatively small unit repeat length of 7.8 kb. There is no heterogeneity in the length of the rDNA repeat units although they display limited differences in either base sequence or pattern of methylation. The cloned rDNA sequences are shown to comprise the entire repeat unit and have been used to obtain a detailed restriction map as well as an approximate transcription map of soybean rRNA genes. The cloning of rDNA from soybean suggests that recombinant DNA techniques can be successfully applied to the genomic DNA of higher plants despite the high degree of methylation exhibited by plant DNA.

摘要

重组DNA载体λ噬菌体Charon 4A被用于构建大豆(Glycine max)基因组DNA的DNA序列文库。为了确定大豆基因组中核糖体DNA(rDNA)的组织方式,已从该文库中分离出含有与17S和25S rRNA互补序列的克隆,并将其与Southern印迹杂交结合使用。rRNA基因以串联方式重复,单位重复长度相对较小,为7.8 kb。rDNA重复单元的长度没有异质性,尽管它们在碱基序列或甲基化模式上存在有限差异。克隆的rDNA序列显示包含整个重复单元,并已用于获得大豆rRNA基因的详细限制性图谱以及大致转录图谱。从大豆中克隆rDNA表明,尽管植物DNA表现出高度甲基化,但重组DNA技术仍可成功应用于高等植物的基因组DNA。

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