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  • Title: [Construction and identification of yeast expression vectors containing human platelet-derived growth factor B-chain gene].
    Author: Yang CL, Chen JT, Deng F, Tang HZ, Tan XY, Luo SQ.
    Journal: Di Yi Jun Yi Da Xue Xue Bao; 2004 Mar; 24(3):273-6. PubMed ID: 15041539.
    Abstract:
    OBJECTIVE: To investigate the expression efficiency of human platelet-derived growth factor B-chain (PDGF-B) gene in yeast and assess the activity of the expressed product. METHODS: A full-length complementary DNA of human PDGF-B gene was amplified from the total RNA extracted from human vascular endothelial cells using reverse transcription (RT)-PCR and then cloned into pGEM-T vector. The PCR products with specific primers were recombined into yeast expression plasmids pMETB or pMETalphaA, followed by identification with restriction endonuclease. RESULTS: The 578 bp fragment encoding mature PDGF-BB peptide with signal peptide and the 340 bp fragment without signal peptide were identified and verified by restriction endonuclease mapping and sequencing, and two yeast expression vectors pMETB-PDGFB(1) and pMETalphaA-PDGFB(2) were reconstructed. CONCLUSION: The yeast expression vectors containing human PDGF-B gene have been successfully constructed for further studies of gene expression in yeast.
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