A SACCHAROMYCES-CEREVISIAE GENOMIC PLASMID BANK BASED ON A CENTROMERE-CONTAINING SHUTTLE VECTOR
A SACCHAROMYCES-CEREVISIAE GENOMIC PLASMID BANK BASED ON A CENTROMERE-CONTAINING SHUTTLE VECTOR
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DOI：
10.1016/0378-1119(87)90232-0
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发表时间：
1987-01-01
期刊：
影响因子：
3.5
通讯作者：
FINK, GR
中科院分区：
文献类型：
--
作者：
ROSE, MD;NOVICK, P;FINK, GR
利用含着丝粒的酵母穿梭载体YCp50构建了一套基因组质粒库。含有着丝粒的载体可用于分离高拷贝数时对酵母有毒的基因。准备了14个独立的组，每个组的平均代表是酵母基因组的两到三倍。来自给定银行的任何单独的质粒都被保证与从每个其他银行获得的质粒具有独立的起源。这些库是由三种不同大小的DNA片段组成的，这些DNA片段是由不同条件的Sau3A部分消化产生的。这避免了由于位点对Sau3A切割的不同敏感性而引起的偏差。插入的DNA足够大，以至于大多数基因将以大约0.1%的频率出现在这组质粒库中。
A set of genomic plasmid banks was constructed using the centromere-containing yeast shuttle vector YCp50. The centromere-containing vector is useful for the isolation of genes that are toxic to yeast when present in high copy number. Fourteen independent banks were prepared each with an average representation of two to three times the yeast genome. Any individual plasmid from a given bank is guaranteed to be of independent origin from plasmids obtained from each of the other banks. The banks were constructed from three different size classes of DNA fragments that resulted from varying conditions of partial digestion with Sau3A. This avoided the bias caused by differential sensitivity of sites to cleavage with Sau3A. Insert DNA is sufficiently large that most genes will be present in the set of plasmid banks at a frequency of about 0.1%.