同源重组
遗传学
生物
体外重组
酵母人工染色体
基因组
克隆(编程)
FLP-FRT重组
DNA
基因
插入(复合材料)
分子克隆
计算生物学
遗传重组
重组
基因定位
染色体
计算机科学
肽序列
程序设计语言
机械工程
工程类
作者
Forrest Spencer,Gary Ketner,Carla Connelly,Phil Hieter
出处
期刊:Methods
[Elsevier]
日期:1993-08-01
卷期号:5 (2): 161-175
被引量:43
标识
DOI:10.1006/meth.1993.1021
摘要
YAC cloning technology has contributed significantly to physical mapping of genomes and holds enormous potential for identification and functional analysis of genes on large DNA segments. This potential is greatly enhanced by the ability to use homologous recombination-based methodologies to restructure or manipulate exogenous DNA in the yeast host. Three methods are described for manipulation of YAC DNA. "Recombinationally targeted YAC cloning" involves the capture of large DNA segments by recombination in vivo between the YAC vector arms and a target DNA segment. "Two-step gene replacement" exploits targeted homologous recombination to replace a selected segment within a YAC with mutant or exogenous sequences. "YAC transfer by kar1 mating" allows rapid movement of YACs under study into yeast genetic backgrounds that facilitate homologous recombination-based manipulation. These three methods taken together will aid significantly in the construction of physical genome maps and provide powerful genetic tools for the study of genome function.
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