中国农业科学2012，Vol.45Issue(5)：999-1009,11.DOI:10.3864/j.issn.0578-1752.2012.05.022
鹅β-防御素10基因的分离、鉴定与表达
Isolation, Characterization, and Expression of Goose Avian β-Defensin 10
摘要
Abstract
[Objective] The objective of this study was to clone avian p-defensin 10 (AvBDIO) gene from goose tissues and to characterize the antimicrobial activity of the recombinant GST-AvBD 10 fusion protein. [ Method ] The coding sequences of goose AvBDIO was obtained from kidney of goose by RT-PCR, phylogenetic relationships of the goose AvBDIO with those of other avian species and some mammalian were analyzed. In addition, tissue distribution of the gene was detected by real time PCR. AvBD gene coding sequence was inserted into the pGEX-6p-l vector. The constructs were transformed into competent Escherichia coli BL21 (DE3) cells. Expression of the fusion proteins was induced with isopropyl P-D-1-thiogalactopyranoside (IPTG) and the proteins were purified. Furthermore, bioactivities of the recombinant protein were analyzed in vitro. [Result] The full length cDNA of goose AvBDIO consisted of 207 nucleotide acids, encoding 68 amino acid residues. Phylogenetic analysis demonstrated that the goose AvBDIO shared the highest amino acid homology with duck AvBDIO (92.7%). The recombinant protein exhibited high antimicrobial activity against 12 bacterial strains investigated. The antimicrobial activity was decreased in high salt concentration and exhibited no hemolytic properties. {Conclusion] The goose AvBDIO gene from goose was successfully cloned and expressed in E. coli. The purified recombinant protein showed antimicrobial activity and no hemolytic properties.关键词
鹅AvBD10/克隆/遗传进化分析/组织表达/抗菌活性Key words
goose AvBDIO/ clone/ phylogenetic analysis/ tissue distribution/ antimicrobial activity引用本文复制引用
周财源,张名岳,韩宗玺,邵昱昊,刘胜旺,马得莹..鹅β-防御素10基因的分离、鉴定与表达[J].中国农业科学,2012,45(5):999-1009,11.基金项目
国家自然科学基金(30972110)、黑龙江省教育厅科研重点项目(11531z05) （30972110）
