The complete set of predicted genes from Saccharomyces cerevisiae in a readily usable form

被引:98
作者
Hudson, JR
Dawson, EP
Rushing, KL
Jackson, CH
Lockshon, D
Conover, D
Lanciault, C
Harris, JR
Simmons, SJ
Rothstein, R
Fields, S [1 ]
机构
[1] Univ Washington, Dept Genet, Seattle, WA 98195 USA
[2] Res Genet Inc, Huntsville, AL 35801 USA
[3] BioVentures Inc, Mufreesboro, TN 37129 USA
[4] Univ Washington, Dept Med, Seattle, WA 98195 USA
[5] Columbia Univ, Dept Genet & Dev, New York, NY 10032 USA
来源
GENOME RESEARCH | 1997年 / 7卷 / 12期
关键词
D O I
10.1101/gr.7.12.1169
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Nearly all of the open reading frames (ORFs) of the yeast Saccharomyces cerevisiae have been synthesized by PCR using a set of similar to 6000 primer pairs. Each of the forward primers has a common 22-base sequence at its 5' end, and each of the back primers has a common 20-base sequence at its 5' end. These common termini allow reamplification of the entire set of original PCR products using a single pair of longer primers-in our case, 70 bases. The resulting 70-base elements that flank each ORF can be used for rapid and efficient cloning into a linearized yeast vector that contains these same elements at its termini. This cloning by genetic recombination obviates the need for ligations or bacterial manipulations and should permit convenient global approaches to gene function that require the assay of each putative yeast gene.
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页码:1169 / 1173
页数:5
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