人β-防御素4基因在COS-7细胞中的转染表达  

Eukaryotic expression of human β defensin 4 gene in COS-7 cells

作　　者：曹玉红[1] 张光运[2] 成胜权[1] 李如英[1] 丁翠玲[1] 

机构地区：[1]第四军医大学西京医院儿科,陕西西安710032 [2]第四军医大学西京医院神经内科,陕西西安710032

出　　处：《西部医学》2012年第9期1668-1670,共3页Medical Journal of West China

基　　金：陕西省科技攻关项目(编号:2009K12-01)

摘　　要：目的探讨人β-防御素4(HBD4)基因在真核细胞COS-7中表达的可能性。方法用脂质体转染法将实验室已构建的重组真核表达载体pcDNA3.1+/HBD4导入COS-7细胞,采用RT-PCR、Western-blot法检测HBD4的mRNA和蛋白表达水平,并对表达的HBD4进行抗菌活性的初步研究。结果 pcDNA3.1+/HBD4转染的COS-7细胞中检测到HBD4mRNA表达。Western blot分析显示,在pcDNA3.1+/HBD4转染的COS-7细胞及细胞培养液中检测到了HBD4蛋白的表达。Kirby-Bauer纸片扩散法证实表达的HBD4蛋白对大肠杆菌ATCC25922具有较强的杀伤作用。结论在COS-7细胞中成功表达具有抗菌活性的HBD4多肽。Objective To study the possibility of HBD4 gene expression in eukaryotic cells COS-7. Methods The recombinant expression vector pcDNA3.1 ＋/HBD4 was transfected into COS-7 cells with liposome transirection protocol. RT-PCR and Western blot were used to identify the expressions of HBD4 mRNA and HBD4 peptide. Antimicrobial activity of the secreted HBD4 was evaluated. Results HBD4 mRNA expression was detected by RT-PCR in the transfected cells. Western blot analysis showed that HBD4 expressed in both transfected cells and cell culture fluid. That HBD4 protein secreted into the culture medium showed strong antimicrobial activity against E. coli ATCC25922. Conclusions COS- 7 cells transfected with pcDNA3. I＋/HBD4 can express bioactive HBD4.

关 键 词：人β-防御素4 真核表达 基因转染 

分 类 号：Q786[生物学—分子生物学]