The Cloning Of Human Neutrophil Defensin-1(HNP-1) Gene And Construction Of Its Expression Vector

Defensins, a family of small cationic peptide with three to four intramolecular cysteine disulfide bonds, have a broad spectrum of antimicrobial activity against bacterial, fungiand even some enveloped viruses. As multifunctional effector molecules of innate immunity, defensins are the key components in immune system. In this report, a pair of primers were designed and synthesized according to published l-INP-1 eDNA nucleotide sequence. Total HL-60 cells RNA was isolated from cultured I-IL-60 cells. Then the target fragment was amplified by RT-PCR and cloned into pUC 18 vector between two restriction enzyme sites(BamH I and Xal I). The sequence analysis showed the cloned fragment was I-INP-l eDNA. Furthermore, the recombinant was digested by Sma I and Sal I and inserted into pGEX-6p- 1 expression vector. After screening by PCR and restriction analysisi, we concluded HNP-l expression vector was constructed. The achievement of this study established the foundation for the further research on expressing I-INP- 1. Postgraduate: Hua Ling Major: Microbiology and Immunology Tutor: Prof. Jingpeng Li...