小鼠BPI N端功能基因片段的克隆及其表达产物在
lonning of N-terminal functional gene fragment of murine BPI gene and studies on its intracellular bacteriocide function
目的：克隆小鼠 BPI36-259基因片段，转染细胞获得具有杀菌作用的目的抗菌蛋白。方法：采用生物信息学方法，构建PUC57-muBPI1-281质粒；PCR扩增获得muBPI36-259基因片段，制备pcDNA3.1 (+)-muBPI36-259重组质粒；采用电穿孔法将上述重组质粒转染RAW264.7细胞，Western-blot法检测目的蛋白在胞内的表达；建立胞内杀菌实验模型，检测muBPI36-259目的蛋白对胞内寄生菌（伤寒杆菌）的杀伤作用。结果：muBPI36-259目的蛋白在RAW264.7细胞内成功表达，对胞内寄生菌—G-伤寒杆菌具有杀伤作用。结论：成功克隆了小鼠BPI36-259基因片段，转染RAW264.7细胞获得具有生物学活性目的抗菌蛋白。
Objective: Clone the N-terminal fragment of mouse BPI from 36 to 259 amino acid, get the target protein with bacteriocide function by transfected to RAW264.7 cells. Methods: We synthesized PUC57-muBPI1-281 plasmid by comparision to human BPI N-terminal functional fragment, then got mouse BPI36-259 functional fragment by PCR, this fragment was cloned in expression vector pcDNA3.1 (+) to construct pcDNA3.1 (+)-muBPI36-259 recombinant plasmid; by transfected to RAW264.7 cells using electroporation method, target protein expression was detected by Western-blot method. Furthermore, bacteriocide function to intracellular Salmonella typhi was found on this recombinant protein. Results: The target protein of muBPI36-259 expressed in RAW264.7 cells successfully, and it could kill intracellular bacteriocide—G- Salmonella typhi. Conclusion: We has successfully cloned mouse BPI36-259 gene fragment, transfected into RAW264.7cells and got the target antibacterial protein of muBPI36-259 .
刘振龙、高燕、吕喆、王炜、冯颖、孔庆利、安云庆、李丽、赵冬
分子生物学细胞生物学生物化学
杀菌/渗透性增强蛋白RAW264.7细胞伤寒杆菌
bactericidal/permeability-increasing proteinRAW264.7cellSalmonella typhi
刘振龙,高燕,吕喆,王炜,冯颖,孔庆利,安云庆,李丽,赵冬.小鼠BPI N端功能基因片段的克隆及其表达产物在[EB/OL].(2008-04-10)[2025-07-16].http://www.paper.edu.cn/releasepaper/content/200804-328.点此复制
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