Use of gap repair in fission yeast to obtain novel alleles of specific genes.

Abstract:

:We have adapted a method for making libraries of mutations in any specific gene for use in the fission yeast Schizosaccharomyces pombe . This elegant and simple method consists of PCR amplification of the gene of interest, followed by co-transformation of fission yeast with the PCR fragment and a linearized plasmid vector prepared such that the ends of the vector share DNA sequence with the ends of the PCR fragment. Homologous recombination between the vector and the PCR fragment occurs at a high frequency and results in a collection of yeast transformants, most harboring a mutated allele of the original gene within the vector of choice. This library can then be screened or selected for phenotypes of interest.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Kostrub CF,Lei EP,Enoch T

doi

10.1093/nar/26.20.4783

subject

Has Abstract

pub_date

1998-10-15 00:00:00

pages

4783-4

issue

20

eissn

0305-1048

issn

1362-4962

pii

gkb741

journal_volume

26

pub_type

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