RRC ID 28675
Author Matsuyama A, Shirai A, Yashiroda Y, Kamata A, Horinouchi S, Yoshida M.
Title pDUAL, a multipurpose, multicopy vector capable of chromosomal integration in fission yeast.
Journal Yeast
Abstract A novel series of plasmid vectors named pDUAL have been developed. These vectors enable one to introduce not only multicopies of genes with episomal maintenance but also a single copy with chromosomal integration into the fission yeast, Schizosaccharomyces pombe. The multicopy plasmids can be easily converted to fragments for chromosomal integration by digestion of the plasmids with a certain restriction endonuclease before transformation of the yeast cells. The resultant fragments, lacking the autonomously replicating sequence, are designed for targeting into the chromosomal leu1 locus by homologous recombination. Whether the transformants are the results of episomal maintenance of the plasmid or homologous gene targeting can be readily checked by their requirement for uracil or leucine, or by the PCR diagnostic analysis. Furthermore, we propose the use of pDUAL derivatives for PCR-based chromosomal tagging of a gene to introduce several tags into 5'-terminus of a gene, employing a set of primers. Using these all-in-one vectors, a suitable mode of expression of a cloned gene can be selected for individual analysis without any complicated subcloning processes.
Volume 21(15)
Pages 1289-305
Published 2004-11
DOI 10.1002/yea.1181
PMID 15546162
MeSH Base Sequence Chromosomes, Fungal / genetics* Chromosomes, Fungal / metabolism Cloning, Molecular Genes, Fungal / genetics Genes, Fungal / physiology Genetic Vectors / genetics* Genetic Vectors / metabolism Green Fluorescent Proteins / genetics Green Fluorescent Proteins / metabolism Microscopy, Fluorescence Molecular Sequence Data Plasmids / genetics* Plasmids / metabolism Polymerase Chain Reaction Recombination, Genetic / physiology* Schizosaccharomyces / genetics* Schizosaccharomyces / metabolism Sequence Analysis, DNA Transformation, Genetic / physiology
IF 2.283
Times Cited 59
WOS Category MYCOLOGY BIOTECHNOLOGY & APPLIED MICROBIOLOGY MICROBIOLOGY BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
DNA material pDUAL (RDB06178) pDUAL2 (RDB06192) pDUAL-HFF1c (RDB06130) pDUAL-HFF41c (RDB06131) pDUAL-HFF81c (RDB06132) pDUAL-HFG1c (RDB06133) pDUAL-HFG41c (RDB06134) pDUAL-HFG81c (RDB06135) pDUAL-FFH1c (RDB06142) pDUAL-FFH41c (RDB06143) pDUAL-FFH81c (RDB06144) pDUAL-GFH1c (RDB06145) pDUAL-GFH41c (RDB06146) pDUAL-GFH81c (RDB06147) pDUAL2-HFF1c (RDB06154) pDUAL2-HFF41c (RDB06155) pDUAL2-HFF81c (RDB06156) pDUAL2-HFG1c (RDB06157) pDUAL2-HFG41c (RDB06158) pDUAL2-HFG81c (RDB06159) pDUAL2-FFH1c (RDB06166) pDUAL2-FFH41c (RDB06167) pDUAL2-FFH81c (RDB06168) pDUAL2-GFH1c (RDB06169) pDUAL2-GFH41c (RDB06170) pDUAL2-GFH81c (RDB06171) pDUAL-HFF1 (RDB06179) pDUAL-HFF41 (RDB06180) pDUAL-HFF81 (RDB06181) pDUAL-HFG1 (RDB06182) pDUAL-HFG41 (RDB06183) pDUAL-HFG81 (RDB06184) pDUAL-FFH1 (RDB06185) pDUAL-FFH41 (RDB06186) pDUAL-FFH81 (RDB06187) pDUAL-GFH1 (RDB06188) pDUAL-GFH41 (RDB06189) pDUAL-GFH81 (RDB06190) pDUAL2-HFF1 (RDB06193) pDUAL2-HFF41 (RDB06194) pDUAL2-HFF81 (RDB06195) pDUAL2-HFG1 (RDB06196) pDUAL2-HFG41 (RDB06197) pDUAL2-HFG81 (RDB06198) pDUAL2-FFH1 (RDB06199) pDUAL2-FFH41 (RDB06200) pDUAL2-FFH81 (RDB06201) pDUAL2-GFH1 (RDB06202) pDUAL2-GFH41 (RDB06203) pDUAL2-GFH81 (RDB06204) pHFF1 (RDB06580) pHFF41 (RDB06581) pHFF81 (RDB06582) pHFG1 (RDB06583) pHFG41 (RDB06584) pHFG81 (RDB06585) pFFH1 (RDB06586) pFFH41 (RDB06587) pFFH81 (RDB06588) pGFH1 (RDB06589) pGFH41 (RDB06590) pGFH81 (RDB06591).