Table I

Plasmids Used in This Study

PlasmidParent vectorRelevant markers and construction*Reference or source
pBluescriptII KS (+)ampr f1 originStratagene
pGEX-2TPtac with gene encoding GSTPharmacia
pQE30Ptac with 6XHis polylinkerQiagen
pRS306URA3 f1 origin(Sikorski and Hieter, 1989)
pRS315CEN6 ARSH4 LEU2 f1 origin(Sikorski and Hieter, 1989)
pRS316CEN6 ARSH4 URA3 f1 origin(Sikorski and Hieter, 1989)
pGF27pRS3042-μm origin (YEp24 fragment) inserted at AatII siteG. Zhu
pGF29pRS3062-μm origin (YEp24 fragment) inserted at AatII siteG. Zhu
pJG28pSB5ampr trc promoter, CMD1J. Geiser
pJP10-2BYCp50URA3 MYO2 CEN4 ARS1(Johnston et al., 1991)
pLI831pBluescriptII SK (+)ADE3E. Muller
myo4Δ::URA3pBluescriptII KS (+)myo4Δ::URA3S. Brown
pRS23pBluescriptII SK (+)5.6-kb ClaI fragment of MYO2This study
pRS28pRS27myo2Δ::TRP1This study
pRS29pRS255.6-kb ClaI fragment of MYO2This study
pRS31pGF295.6-kb ClaI fragment of MYO2This study
pRS43pRS29NheI site removedThis study
pRS50pRS372-μm origin (YEp24 fragment) inserted at NotI siteThis study
pRS72pRS43NcoI site added at codon 1 of MYO2This study
pRS172pRS43Internal deletion of MYO2 (S787GThis study
 Δaa788–927) also known as MYO2-D6IQ
pRS174pRS3065.4-kb ClaI fragment of MYO2-D6IQThis study
pRS221pGF295.4-kb ClaI fragment of MYO2-D6IQThis study
pRS276pBluescriptII KS (+)3.3-kb BamHI fragment carrying MLC1This study
pRS286pRS285mlc1Δ::TRP1This study
pRS289pRS3161.4-kb BamHI-SacI fragment of MLC1This study
pRS290pGF271.4-kb BamHI-SacI fragment of MLC1This study
pRS296pQE30ampr promoter 6XHis-MLC1This study
pRS321pRS3151.4-kb BamHI-SacI fragment of MLC1This study
pSB6pACYC177kanr lysis genes of lambda(Brockerhoff et al., 1994)
pSB20pGEX-3XMYO2 (aa740–1457) fused to GST(Brockerhoff et al., 1994)
pSB21pGEX-3XMYO2 (aa740–1116) fused to GST(Brockerhoff et al., 1994)
pSB24pGEX-3XMYO2 (aa790–924) fused to GST(Brockerhoff et al., 1994)
pSB25pGEX-3XMYO2 (aa740–833) fused to GST(Brockerhoff et al., 1994)
pSB27pGEX-3XMYO2 (aa247–740) fused to GST(Brockerhoff et al., 1994)
pTD28pTD172-μm origin LYS2 CMD1(Davis and Thorner, 1989)
pTD29YEp242-μm origin LYS2(Geiser et al., 1993)
  • *  Unless otherwise stated, all markers from the parent plasmid are present in the new construct. MYO2-Δ6IQ encodes an internal deletion in Myo2p with the mutation S787G and missing amino acids 787–927. MLC1 is identical to the Saccharomyces cerevisiae open reading frame YGL106w.