利用重复序列指纹图谱进行人类基因组物理图谱绘制的一个测试案例：构建亚克隆到黏粒中的420 kb酵母人工染色体的物理图谱。

A test case for physical mapping of human genome by repetitive sequence fingerprints: construction of a physical map of a 420 kb YAC subcloned into cosmids.

作者信息

Bellanné-Chantelot C, Barillot E, Lacroix B, Le Paslier D, Cohen D

机构信息

Centre d'Etude du Polymorphisme Humain, Paris, France.

出版信息

Nucleic Acids Res. 1991 Feb 11;19(3):505-10. doi: 10.1093/nar/19.3.505.

DOI:10.1093/nar/19.3.505

PMID:2011524

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC333640/

Abstract

A rapid and safe method of Yeast Artificial Chromosome (YAC) physical mapping by cosmid 'fingerprinting' is presented. YACs are subcloned into cosmids which are prepared without previous separation of cloned DNA from host DNA. Groups of overlapping clones are detected according to their restriction fragments size and intensity after hybridization with total human DNA. To test this approach, a cosmid library was constructed from total DNA of a yeast strain containing a 420 kb YAC. A single contig of 84 clones was obtained with a minimal detectable overlap of 60% i.e. a 9.2 fold representative library. Large scale physical mapping of YACs would take full advantage of the DNA preparation procedure employed in this work and allows to take into account restriction fragment intensities.

摘要

本文介绍了一种通过黏粒“指纹分析”进行酵母人工染色体（YAC）物理图谱构建的快速且安全的方法。将YAC亚克隆到黏粒中，制备黏粒时无需事先从宿主DNA中分离克隆DNA。与总人类DNA杂交后，根据重叠克隆组的限制性片段大小和强度来检测它们。为测试该方法，从含有一个420 kb YAC的酵母菌株的总DNA构建了一个黏粒文库。获得了一个由84个克隆组成的单一重叠群，最小可检测重叠率为60%，即一个9.2倍代表性文库。YAC的大规模物理图谱构建将充分利用本研究中采用的DNA制备程序，并能够考虑限制性片段强度。