中国组织工程研究与临床康复2011，Vol.15Issue(24)：4549-4552,4.DOI:10.3969/j.issn.1673-8225.2011.24.044
抗菌肽Cecropin A基因原核表达及表达产物的鉴定
Prokaryotic expression of antibiotic peptide Cecropin A gene and identification of expression products
陈磊1
作者信息
- 1. 河南中医学院第一附属医院心内科,河南省郑州市,450000
- 折叠
摘要
Abstract
BACKGROUND: Cecropins are a kind of micromolecule protein with antibacterial activity. Eukaryotic cell-expressed or artificially synthesized Cecropins is characterized by low efficiency and high cost. OBJECTIVE: To clone and express an antibacterial peptide gene of Musca domestica Cecropin A, and to identify recombinant expression product. METHODS: Mature Musca domestica Cecropin A encoding nucleotide sequence was searched from the GenBank and amplified by RT-PCR. The gene of Musca domestica Cecropin A was cloned into prokaryotic expression vector pET32a and fused with gene of Thioredoxin (Trx) and expressed in E.coli BL2l (DE3). After induction by isopropyl-β-D-thiogalactoside, the sera of the immunized rabbits were collected after rabbits were immunized with the hemolymph of housefly larvae. Recombinant protein was identified by western blot analysis and N-[Tris(hydroxymethyl)methyl]glycine-sodium dodecylsulfate-polyacrylamide gel electrophoresis.RESULTS AND CONCLUSION: After induction by isopropyl-β-D-thiogalactoside, E.coli BL21 expressed mature Cecropin. Rabbit anti- housefly larvae sera, N-[Tris(hydroxymethyl)methyl]glycine-sodium dodecylsulfate-polyacrylamide gel electrophoresis and western blot analysis results confirmed that expression products were mature Cecropin. These suggest that prokaryotic expression system can be utilized to obtain natural mature Cecropin.关键词
家蝇/Cecropin A/多克隆抗体/融合表达/三羟甲基氨基甘氨酸-十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分类
基础医学引用本文复制引用
陈磊..抗菌肽Cecropin A基因原核表达及表达产物的鉴定[J].中国组织工程研究与临床康复,2011,15(24):4549-4552,4.
