JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES)

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Construction of pEGFPKallikrein expression vectorand its expression in endothelial progenitor cells in rabbit

JIANG Hong,LI Da-qing.ZHANG Yun,LIU Chun-xi,FENG Jin-bo,WANG Rong   

  1. Department of Cardiology, Qilu Hospital of Shandong University, Jinan 250012, Shandong, China
  • Received:2006-03-29 Revised:1900-01-01 Online:2006-06-24 Published:2006-06-24
  • Contact: JIANG Hong

Abstract: To construct pEGFPKallikrein eukaryotic expression vector by using green fluorescence protein as reportor gene and transfecting rabbit endothelial progenitor cells. Methods： The human tissue kallikrein（tHK） genes were amplified with PCR technique and inserted into pEGFPN3 vector. The rabbit endothelial progenitor cells (EPCs) were transfected with the formed plasmid by means of lipidosome. The KallikreinEGFP fused protein was viewed directly with fluorensce microscope, and the expression of tHK was detected by RTPCR(reverse transcriptionPCR) and ELISA. Results： Plasmids was formed correctly and was detected by PCR meanwhile it being sequenced and expressed in the rabbit EPCs. The fused protein has activites of both kallikrein and EGFP. Conclusion: tHK gene modified EPCs could play a role in the treatment of artery injury and endothelial dysfunction.

Key words: Kallireins, Endothelium, Stem Cells, Gene Transfer Techniques

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