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. 2009 Feb 27;284(9):5602–5609. doi: 10.1074/jbc.M806664200

TABLE 1.

Plasmids and yeast strains used in this study

Plasmid (description) Comments
PEC2036 (AD-vector) pGADT7-Rec from Clontech TRP1 Ampr
PEC2037 (DBD-vector) pGBKT7 from Clontech LEU2 Kanr
PEC2038 (DBD-proRTD1a) proRTD1a cloned in the NcoI/BamHI sites of pGBKT7
PEC2069 (AD-SDF2L1) SDF2L1/pGADT7-Rec; plasmid expressing interactor of proRTD1a, isolated from yeast
PEC2073 (DBD-proseg) The prosegment of RTD1a cloned in the EcoRI/BamHI sites of pGBKT7
PEC2082 (DBD-proHNP3) PEC2080 (proHNP3/pCR2.1) was digested with Acc65I, filled-in with Klenow and then digested with EcoRI. The fragment containing proHNP3 was then ligated into EcoRI/SmaI-digested pGBKT7
PEC2084 (AD-ΔN28SDF2L1) ΔN28SDF2L1 cloned into EcoRI and XhoI sites of pGADT7-Rec
PEC2105 (AD-ΔN28SDF2L1ΔMIR1) SDF2L1ΔMIR1 (N-terminal deletion, ΔN87) cloned into EcoRI and XhoI sites of pGADT7-Rec
PEC2106 (AD-SDF2L1ΔMIR3) SDF2L1ΔMIR3 (C-terminal deletion, ΔC151) cloned into EcoRI and XhoI sites of pGADT7-Rec
PEC2113 (DBD-proHBD1) proHBD1 cloned as EcoRI/BamHI fragment into pGBKT7
PEC2114 (DBD-proHD5) proHD5 cloned as EcoRI/BamHI fragment into pGBKT7
PEC2122 (DBD-ΔN28SDF2L1ΔMIR2) ΔN28SDF2L1ΔMIR2 (internal deletion of residues 95-149) cloned into the EcoR1/XhoI sites of pGADT7-Rec
PEC2123 (DBD-proHNP3Δ35-49) proHNP3Δ35-49 fragment cloned into EcoRI/BamHI sites of pGBKT7
PEC2124 (DBD-proRTD1aΔRLL) proRTD1aΔRLL cloned into NcoI and BamHI sites of pGBKT7
PEC2125 (DBD-RTD1a-9mer) RTD1a was cloned into the EcoRI and XhoI sites of pGBKT7
PEC2126 (DBD-HNP3) HNP3 fragment was cloned into EcoRI and XhoI sites of pGBKT7
PEC2127 (AD-MIR3) MIR3 fragment (residues 151-221 of SDF2L1) was cloned into EcoRI and XhoI sites of pGADT7-Rec
PEC2003 (GST) NcoI site in PEC615 (UAF30/pCBGST1) was filled in with Klenow and re-ligated to render the UAF30 sequence out of frame, for expression of GST TRP1 Ampr
PEC2007 (GST-proRTD1a) proRTD1a cloned in the NcoI/Acc65I sites of pCBGST1 for expression of GST-proRTD1a
PEC2075 (GST-ΔN28SDF2L1) ΔN28SDF2L1 cloned into the NcoI/Acc65I sites of pCBGST1 for expression of GST-ΔN28SDF2L1
PEC2121 (GST-proHNP3) proHNP3 cloned into the NcoI/Acc65I sites of pCBGST1 for expression of GST-proHNP3
Yeast strains Phenotype
AH109 MATa trp1-901 leu2-3, 112 ura3-52 his3-Δ200 gal4Δ gal80Δ LYS2::GAL1UAS-GAL1TATA-HIS3 GAL2UAS-GAL2TATA-ADE2 URA3::MEL1UAS-MEL1TATA-lacZ MEL1
JD47-13C MATa his3-Δ200 trp1-Δ63 lys2-801 ura3-52 leu2-3, 112