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严格控制质粒的迭代子与DNA指纹分析

Iterons of stringently controlled plasmids and DNA fingerprinting.

作者信息

Huebscher K J, Dolf G, Frey J

机构信息

Institute of Animal Breeding, University of Berne, Switzerland.

出版信息

EXS. 1993;67:79-85. doi: 10.1007/978-3-0348-8583-6_8.

Abstract

DNA probes which detect polymorphic, repetitive sequences in a variety of genomes have been developed using different approaches. Naturally occurring plasmids, with repeated units termed iterons near or within their origins (ori) or replication, could be of interest for the development of probes, possibly even revealing novel minisatellite families in mammals involved in replicational processes. We used the plasmids P1, pSC101 and RSF1010 or their PCR amplified ori regions as probes in Southern blot hybridisations with mammalian DNA. At low stringency they generated reproducible fingerprint-like patterns. A bovine genomic library was screened at the same stringency with the PCR-amplified ori region of P1 containing the five times repeated core sequence 5'-ATGTGTGNTGNNGGG-3' to generate a probe for cattle DNA with higher specificity.

摘要

利用不同方法已开发出能检测多种基因组中多态性重复序列的DNA探针。天然存在的质粒,在其复制起点(ori)附近或内部具有称为迭代子的重复单元,可能对探针开发有意义,甚至可能揭示参与复制过程的哺乳动物中新的小卫星家族。我们将质粒P1、pSC101和RSF1010或它们经PCR扩增的ori区域用作探针,与哺乳动物DNA进行Southern印迹杂交。在低严谨度下,它们产生了可重复的指纹状图谱。用含有五次重复核心序列5'-ATGTGTGNTGNNGGG-3'的P1的PCR扩增ori区域,在相同严谨度下筛选牛基因组文库,以生成特异性更高的牛DNA探针。

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