Construction of eukaryotic expression vector encoding antimicrobial peptides PR39 and its expression and antimicrobial function in RAW264.7 cells

文阳安; 郭金英; 余晓林; 岑东; 陈彦; 张键; 罗淼; 涂植光

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Construction of eukaryotic expression vector encoding antimicrobial peptides PR39 and its expression and antimicrobial function in RAW264.7 cells

更新时间：2026-01-09

- Construction of eukaryotic expression vector encoding antimicrobial peptides PR39 and its expression and antimicrobial function in RAW264.7 cells
- Journal of Army Medical University Vol. 30, Issue 21, Pages: 2044-2046(2008)
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  CLC： R392.11;R394-33;R978.16
- Published：2008
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文阳安,郭金英,余晓林,岑东,陈彦,张键,罗淼,涂植光.抗菌肽PR39真核表达载体的构建及其在巨噬细胞中的表达和抗菌作用[J].陆军军医大学学报(原第三军医大学学报),2008,30(21):2044-2046.

　WEN Yang-an,GUO Jin-ying,YU Xiao-lin,CEN Dong,CHEN Yan,ZHANG Jian,LUO Miao,TU Zhi-guang.Construction of eukaryotic expression vector encoding antimicrobial peptides PR39 and its expression and antimicrobial function in RAW264.7 cells[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(21):2044-2046.
文阳安,郭金英,余晓林,岑东,陈彦,张键,罗淼,涂植光.抗菌肽PR39真核表达载体的构建及其在巨噬细胞中的表达和抗菌作用[J].陆军军医大学学报(原第三军医大学学报),2008,30(21):2044-2046. DOI：

　WEN Yang-an,GUO Jin-ying,YU Xiao-lin,CEN Dong,CHEN Yan,ZHANG Jian,LUO Miao,TU Zhi-guang.Construction of eukaryotic expression vector encoding antimicrobial peptides PR39 and its expression and antimicrobial function in RAW264.7 cells[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(21):2044-2046. DOI：

摘要

目的构建抗菌肽PR39的真核表达载体，转染后观察PR39在小鼠巨噬细胞RAW264.7中的表达及抗菌作用。方法采用拼接PCR法合成抗菌肽PR39基因，克隆到真核表达载体pIRES2-GFP中，经鉴定后用阳离子脂质体转染小鼠巨噬细胞RAW264.7，通过RT-PCR和免疫荧光法检测目的基因的表达，并对其抗菌活性进行初步检测。结果成功构建了抗菌肽PR39的真核表达载体pIRES2-GFP/PR39，并能在小鼠巨噬细胞RAW264.7中表达抗菌肽PR39。表达抗菌肽PR39的巨噬细胞杀灭和清除胞内菌的能力明显增强。结论抗菌肽PR39基因能够在小鼠巨噬细胞RAW264.7中表达并发挥抗菌作用。

Abstract

Objective To construct an eukaryotic expression vector encoding antimicrobial peptide PR39

and observe its expression and antimicrobial function in mouse macrophage RAW264.7 cells. Methods PR39 gene sequence was synthesized with PCR amplification and inserted into an eukaryotic expression vector pIRES2-GFP. After identification

pIRES2-GFP/PR39 vector was transfected into RAW264.7 cells with cationic liposome. The expression of PR39 was detected by RT-PCR and immunofluorescence method

and its antibacterial activity was also detected. Results The pIRES2-GFP/PR39 vector was successfully constructed

and PR39 was expressed in RAW 264.7 cells with antimicrobial activity. RAW 264.7 cells expressing PR39 showed stronger antibacterial ability. Conclusion Antimicrobial peptide PR39 is expressed and shows antimicrobial function in RAW264.7 cells. The successful construction of the PR39 expressing vector have established a foundation for the gene therapy of antimicrobial peptides against intracellular bacterial infection.

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