Construction and identification of yeast expression vectors containing human platelet-derived growth factor B-chain gene

Abstract

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 pMETαA, 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-PDGFB1 and pMETαA-PDGFB2 were reconstructed. Conclusion The yeast expression vectors containing human PDGF-B gene have been successfully constructed for further studies of gene expression in yeast.

CLC Number:

R392.12

YANG Chun-lu¹, CHEN Jian-ting¹, DENG fan², TANG Huan-zhang¹, TAN Xiao-yun¹, LUO shen-qiu². Construction and identification of yeast expression vectors containing human platelet-derived growth factor B-chain gene[J]. Journal of Southern Medical University, 2004, 24(03): 273-276.

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