畜牧兽医学报2011，Vol.42Issue(9)：1309-1315,7.
鸭β-防御素4cDNA的克隆、表达及其抑菌活性的初步分析
cDNA Cloning, Expression and Initial Characterization of Antibacterial Activity of Duck Avian Beta-defensin 4
摘要
Abstract
The objective of the present study was to clone, express, characterize the antimicrobial activity of duck avian β-defensin (AvBD) 4, and determine its mRNA distribution in tissues of duck. Reverse transcription polymerase chain reaction (RT-PCR) was used to amplify duck AvBDA gene from bursa of Fabricius of duck. Sequence analysis showed that the cDNA of duck AvBDA consisted of 171 bp nucleotide, encoding a polypeptide of 56 amino acids. Homology analysis showed that duck AvBD4 shared high percentage of amino acid homology (73. 2 %) with chicken AvBD4. The cDNA of duck AvBD4 mRNA was sub-cloned into EcoR I and Sal I sites of pGEX-6p-l vector to construct recombinant plasmid pGEX-duck AvBD4. The recombinant plasmid was transformed into E. Colt BL21 and the bacteria were induced with IPTG. It was demonstrated by SDS-PAGE that a 32 kD protein which was equal to duck AvBD4 protein in molecular weight was highly expressed.The purified recombinant duck AvBD4 exhibited extensiveantimicrobial activity against 11 strains of bacteria, including Gram-positive and Gram-negative investigated. In high salt ions conditions, recombinant duck AvBD4 protein remained antibacterial activity against Staphylococcus aureus and Pasteurella multocida. In addition, the recombinant protein of he-molytic activity was extremely low. Additionally, real time PCR was used to determine tissue distribution of the gene. It was showed that AvBD4 mRNA was expressed limitedly in tissues of ducks. It was moderately expressed in the cecal tonsil, spleen, and bursa of Fabricius of ducks.关键词
鸭/AvBD4/重组蛋白/抗菌活性/组织分布Key words
duck/AvBD4/recombinant protein/antibacterial activity/tissue distribution分类
农业科技引用本文复制引用
蔺利娟,韩宗玺,邵昱昊,张可心,刘胜旺,李子瑞,马得莹..鸭β-防御素4cDNA的克隆、表达及其抑菌活性的初步分析[J].畜牧兽医学报,2011,42(9):1309-1315,7.基金项目
国家自然科学基金(30972110) （30972110）
黑龙江省教育厅科研重点项目(11531205) （11531205）
