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人类卫星DNA序列的分子克隆及其在检测DNA多态性中的应用。

Molecular cloning of human satellite DNA sequences and their use in detecting DNA polymorphism.

作者信息

Ehtesham N Z, Talwar G P, Ali A, Hasnain S E

机构信息

National Institute of Immunology, Shaheed Jeet Singh Marg, New Delhi.

出版信息

Indian J Biochem Biophys. 1990 Oct;27(5):275-9.

PMID:2079331
Abstract

A approximately 400 bp HaeIII human genomic satellite DNA band was cloned into pUC18 to construct a partial library. A fragment of bacteriophage M13 containing a sequence homologous to the human minisatellite core was cloned in pUC18 and was used as a probe to isolate a approximately 350 bp human satellite clone (pTRF5.6) from the partial library. Other clones from this library showed a wide variation in terms of size and hybridization to the pTRF5.6 clone. Human DNA from different individuals was digested with restriction enzymes, Southern transferred and probed with TRF5.6. Individual-specific complex pattern of DNA bands was produced. TRF5.6, therefore, could be useful as a probe for detecting genetic polymorphism.

摘要

将一条约400 bp的HaeIII人类基因组卫星DNA条带克隆到pUC18中以构建一个部分文库。将含有与人类小卫星核心序列同源的噬菌体M13片段克隆到pUC18中,并用作探针从部分文库中分离出一个约350 bp的人类卫星克隆(pTRF5.6)。该文库中的其他克隆在大小和与pTRF5.6克隆的杂交方面表现出广泛的差异。用限制性内切酶消化来自不同个体的人类DNA,进行Southern印迹转移并用TRF5.6进行杂交检测。产生了个体特异性的DNA条带复杂模式。因此,TRF5.6可用作检测遗传多态性的探针。

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