畜牧兽医学报2011，Vol.42Issue(11)：1638-1642,5.
驯鹿β-防御素-1(reBD-1)基因的原核表达及蛋白活性的鉴定
Prokaryotic Expression of Reindeer β-defensin-1 (reBD-1) Gene and Bioactivity Identification of the Recombinant Protein
摘要
Abstract
The aim of this experiment was to construct prokaryotic expression vector of reindeer β-defensin-l(reBD-l), induce the expression of reBD-1 fusion protein in E. Coli, and evaluate the bioactivity of the expression products. The prepro-peptide of reBD-1 was amplified by RT-PCR. The mature peptide of encoding reBD-1 was amplified from recombinant cloning vector PMD19T/ reBD-1, and then cloned into pET-32a (+ ), in which reBD-1 fusion protein expression was induced by IPTG in E. Coli BL2KDE3). The expressed product was further cultured and purified for experiment of bacteriostasis in vitro. The results showed that the amplified products of prepro-peptide and mature peptide were 215 and 138 bp, respectively, and the homology of the sequences of the targeted gene and reBD-1 mRNA was up to 100%. The molecular weight of fusion proteins of prepro-peptide and mature peptide were 28 and 24 ku. The agar diffusion method has demonstrated that 0. 08 mg · mL-1 purified mature peptide protein has obvious antimicrobial activity against S. Aureus and E. Coli. From the results we can conclude that the prepro-peptide and mature peptides have high expression level in E. Coli, and the mature peptide has resistance to both Gram negative and Gram positive bacteria.关键词
驯鹿/防御素/基因克隆/蛋白表达/抗菌活性Key words
reindeer/ defensin/ gene cloning/ protein expression/ bacteriostasis分类
农业科技引用本文复制引用
苏丽娜,杨银凤,景岚,曹贵方..驯鹿β-防御素-1(reBD-1)基因的原核表达及蛋白活性的鉴定[J].畜牧兽医学报,2011,42(11):1638-1642,5.基金项目
内蒙古农业大学科技创新团队建设项目(NDPYYTD2010-6) （NDPYYTD2010-6）
国家自然基金项目(31160491) （31160491）
