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目的:建立杀菌/渗透增强蛋白(BPI)N-端193个氨基酸的重组表达质粒。方法:利用RT-PCR的方法从HL-60细胞内扩增出BPI氨基端1~193个氨基酸的基因序列,克隆入T载体。将BPI 193基因片段定向克隆入原核表达载体pET-28 a中,构建重组的原核表达质粒pET-BPI 193,转化大肠杆菌BL 21菌株。结果:从HL-60细胞中扩增得到579 bp的BPI 193基因片段,构建了T-BPI 193亚克隆和PET-BPI 193重组表达质粒。结论:成功构建了pET-BPI 193重组表达质粒。
OBJECTIVE: To establish a recombinant expression plasmid with 193 amino acids N-terminal of bactericidal / osmotic enhanced protein (BPI). Methods: The amino acid sequence of 1 193 amino acids of BPI was amplified from HL-60 cells by RT-PCR and cloned into T vector. The BPI 193 gene fragment was cloned into prokaryotic expression vector pET-28a. The recombinant prokaryotic expression plasmid pET-BPI 193 was constructed and transformed into E.coli BL21 strain. Results: A 579 bp BPI 193 gene fragment was amplified from HL-60 cells and subcloned by T-BPI 193 and PET-BPI 193 recombinant plasmid. Conclusion: The pET-BPI 193 recombinant plasmid was successfully constructed.