RRC ID 47484
Author Matsuyama A, Yoshida M.
Title Heterologous gene expression by chromosomal integration in fission yeast.
Journal Methods Mol. Biol.
Abstract Thanks to the convenience and flexibility of the multicopy plasmid-based approach for heterologous gene expression, this technique has long been used for biological studies, especially in prokaryotes and lower eukaryotes. For better understanding of biological mechanisms, however, there are increasing demands on the experimental technologies enabling fine-tuned expression of introduced heterologous genes or serving conditions that are closer to the physiological conditions. For this purpose, the use of direct tagging of a chromosomal gene has been gradually increasing, although the use conditions of this approach are relatively limited compared to plasmid-based methods. Expression of a cloned gene using chromosomal integration has a property intermediate between multicopy plasmid-based method and direct tagging of an endogenous gene. Here, we describe the principle and methods of introduction of a cloned gene into the targeting loci of the chromosome in fission yeast.
Volume 824
Pages 433-50
Published 2012
DOI 10.1007/978-1-61779-433-9_23
PMID 22160913
MeSH Chromosomes, Fungal / genetics* Cloning, Molecular DNA Primers / genetics Gene Expression Regulation, Fungal / genetics* Genes, Fungal / genetics* Genetic Vectors / genetics Plasmids / genetics* Polymerase Chain Reaction Schizosaccharomyces / genetics Schizosaccharomyces / metabolism* Transformation, Genetic / genetics*
Times Cited 3
WOS Category GENETICS & HEREDITY BIOCHEMICAL RESEARCH METHODS BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
DNA material pDUAL (RDB06178) 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) pDUAL-YFH1 (RDB06191) pDUAL2 (RDB06192) 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) pDUAL-Pef1a (RDB06529) pDUAL-Ptub1 (RDB06530) pDUAL-HFF21 (RDB06531) pDUAL-HFF31 (RDB06532) pDUAL-HFF51 (RDB06533) pDUAL-HFF61 (RDB06534) pDUAL-FFH21 (RDB06535) pDUAL-FFH31 (RDB06536) pDUAL-FFH51 (RDB06537) pDUAL-FFH61 (RDB06538) pDUAL-HFG21 (RDB06539) pDUAL-HFG31 (RDB06540) pDUAL-HFG51 (RDB06541) pDUAL-HFG61 (RDB06542) pDUAL-GFH21 (RDB06543) pDUAL-GFH31 (RDB06544) pDUAL-GFH51 (RDB06545) pDUAL-GFH61 (RDB06546) pDUAL2-HFF21 (RDB06547) pDUAL2-FFH21 (RDB06548) pDUAL2-HFF61 (RDB06549) pDUAL2-FFH61 (RDB06550) pLYS1U (RDB06552) pLYS1K (RDB06553) pLYS1H (RDB06554) pLYS1B (RDB06555) pARG1U (RDB06556) pARG1K (RDB06557) pARG1H (RDB06558) pARG1B (RDB06559) pHIS3U (RDB06560) pHIS3K (RDB06561) pHIS3H (RDB06562) pHIS3B (RDB06563) pHIS3K-Ptub1 (RDB06564) pHIS3U-Pnmt1 (RDB06565) pLYS1U-Ptub1 (RDB06566) pLYS1U-Pnmt1 (RDB06567) pHIS3KF (RDB06568) pHIS3UF (RDB06569) pHIS3HF (RDB06570) pHIS3BF (RDB06571) pLYS1KF (RDB06572) pLYS1UF (RDB06573) pLYS1HF (RDB06574) pLYS1BF (RDB06575) pARG1KF (RDB06576) pARG1UF (RDB06577) pARG1HF (RDB06578) pARG1BF (RDB06579)