动物医学进展2025，Vol.46Issue(11)：38-43,6.
绵羊十二指肠中β-防御素-1原核表达和抑菌效果分析
Prokaryotic Expression and Antibacterial Activity Analysis of the β-defensin(SBD-1)Gene in Sheep Duodenal Tissue
摘要
Abstract
Mammalian defensins have antitumor,broad-spectrum antibacterial,and immunomodulatory effects.However,at present,there are certain difficulties in the large-scale production of defensins,so that they have not been widely used.In this study,the recombinant plasmid of pET-22b-SUMO-SBD-1 was constructed and induced to express β-defensin-1(SBD-1)in sheep.The results showed that the total RNA extracted from the duodenum of Mongolian sheep could be amplified by PCR to obtain the sheep SBD-1 cD-NA gene,and in order to express SBD-1 in large quantities,we used a prokaryotic expression system and added SUMO as the tagging protein.In this study,the prokaryotic expression vector pET-22b and the posi-tive recombinant plasmid pMD19-T-SUMO-SBD-1 were treated with Sal Ⅰ and Not Ⅰ double enzyme di-gestion,respectively.The results showed that about 4 000 bp of pET-22b and about 400 bp of SBD-1 were obtained.Subsequently,the recombinant plasmid pET-22b-SUMO-SBD-1 was transformed into BL21(DE3),and the recombinant plasmid pET-22b-SUMO-SBD-1 was detected,and a single band was found to be a positive recombinant bacteria.The results showed that the expression level and activity of SUMO-SBD-1 fusion protein expressed in E.coli may be increased under low temperature conditions.Western blot showed specific bands,and the results were as expected.In this experiment,sheep defensin was expressed in vitro and its antibacterial effect was studied,which laid a foundation for the further development and utilization of defensin.关键词
绵羊β-防御素/SUMO/原核表达/抗菌肽Key words
Sheep β-defensin/SUMO/Prokaryotic expression/Antimicrobial peptide分类
农业科技引用本文复制引用
王鑫,秦少杰,毛伟,巩志国,王志,曹贵方..绵羊十二指肠中β-防御素-1原核表达和抑菌效果分析[J].动物医学进展,2025,46(11):38-43,6.基金项目
国家自然科学基金项目(3216190016) （3216190016）
