抗菌肽CM4与绿色荧光蛋白融合基因的真核载体构建及表达  

Construction and Expression of Eukaryotic Expression Vector of Antibacterial Peptide CM4 and Green Fluorescent Protein Fusion Gene

作　　者：陈玉清[1] 张双全[1] 焦波[1] 秋雯[1] 张杰[1] 

机构地区：[1]南京师范大学生命科学学院江苏省分子医学生物技术重点实验室,江苏南京210097

出　　处：《生物技术通讯》2008年第1期8-10,共3页Letters in Biotechnology

基　　金：国家自然科学基金项目(30271093)

摘　　要：目的:构建绿色荧光蛋白(GFP)与抗菌肽CM4融合基因的真核表达载体。方法:采用递归PCR(rPCR)将GFP基因与CM4基因通过一段核苷酸片段连接成GFP-CM4融合基因,构建真核表达载体pcDNA3-GFP-CM4,用脂质体介导转染人K562白血病细胞,用MTT检测其活性。结果:融合基因可在K562细胞中表达,抗菌肽CM4的表达可抑制K562细胞的生长。结论:为全面了解抗菌肽的生物学活性及其在基因治疗中的可能作用提供了重要的理论依据。Objective： To construct eukaryotic expression vector of antibacterial peptide CM4 and green fluorescent protein（GFP） fusion protein. Methods： GFP-CM4 fusion gene was synthesized by a recursive PCR strategy, in which the CM4 gene was connected to the GFP gene by a linker. Then the sequence was inserted into expression vectors pcDNA3 resulting pcDNA3-GFP-CM4. The construction was introduced into cultured human myeloid leukemia K562 cells with cationic liposome mediated gene transfection technique, and cell growth inhibition was determined by MTI＂ assay. Results The growth of K562 cells was inhibited by the expression of CM4. Conclusion This will be an important basis for the study of the activity of the CM4 and it may expect useful implications for its further use in gene therapy.

关 键 词：抗菌肽CM4 绿色荧光蛋白 真核表达 K562细胞 

分 类 号：Q78[生物学—分子生物学]