pC4-Fv1E plasmid vector质粒载体 BioVector NTCC质粒载体菌种细胞基因保藏中心

pC4-Fv1E plasmid vector质粒载体

Bait and Prey Constructs The pSEL1 and pMG1 vectors for the transient expression of EpoR-LepRF3 and gp130 fusion proteins, respectively, have been previously described [18]. The pCel1f vector, based on the cDNA5/FRT vector for directed, integrated expression has also been described [20]. All primers used in this study are listed in Appendix 1. The bait plasmid, pSEL-eDHFR was generated by cloning the E. coli DHFR gene into pSEL as a SalI- NotI fragment using pBYK-DHFR [13] as a template. The SalI and NotI restriction sites were created by PCR using primers eDHFR-SalI and eDHFR-NotI. pCel1f-eDHFR was constructed by ligating eDHFR to the SacI and NotI sites of pCel1f. SacI and NotI sites were created by PCR with primers eDHFR-SacI and eDHFR-NotI-2. The CDK2, GSK3β, FKBP12F36V and carbonic anhydrase II (CAII) target plasmids were generated by sub-cloning the coding regions of these respective genes into the EcoRI and NotI sites of pMG1. pC4-Fv1E was used to clone FKBP F36V. The EcoRI and NotI restriction sites were created by PCR using primers CDK2F1 and CDK2R1, GSK3β-RI and GSK3β-NI, EcoRI-FKBP and NotI-FKBP CAII-F and CAII-R. The ABL prey construct was generated by cloning the kinase domain of ABLl (amino acids 246-532) into the EcoRI and NotI sites of pMG. The restriction sites were created by PCR using primers EcoRI-ABL3 and NotI-ABL5R. The retroviral vector, pBG-ABL was constructed by sub-cloning the kinase domain of ABL from pMG-ABL into the retroviral vector pBG-ccdB

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