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Cloning and DNA Sequencing for Unstable Minisatellites DNA Regions in E. coli.  

임선희 (동아대학교 자연과학대학 생물학과)
김재우 (동아대학교병원 임상병리)
김광섭 (동아대학교 자연과학대학 생물학)
정윤희 (동아대학교 자연과학대학 생물학)
윤세련 (동아대학교 자연과학대학 생물학)
배호정 (동아대학교 자연과학대학 생물학)
안태진 (동아대학교 자연과학대학 생물학)
선우양일 (동아대학교 자연과학대학 생물학과)
Publication Information
Korean Journal of Microbiology / v.40, no.2, 2004 , pp. 65-72 More about this Journal
Abstract
Instability of some eukaryotic sequence propagated in prokaryotic hosts is a frequently observed phenomenon. It is well documented that long inverted repeats, AT-rich sequences with structures like Z-DNA are extremely unstable in E. coli. These sequences may either be under-represented or even lost when cloned in E. coli. When we analyzed the polymorphic pattern for several tandom repeat (TR) in human SCKI gene, we found some TR regions were frequently deleted from plasmids and had difficult problem for their sequencing. These regions may result in non-clonability of the DNA sequence. Here we have cloned two difficult TR regions under low temperature and made two library for DNA sequencing using a nebulizer or sonicator. This study will help to determine the unstable genomic elements in complex mammalian genome.
Keywords
high GC region; minisatellites; unclonable DNA;
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