Eukaryotic Expression and Purification of Mature Bovine Neutrophil β-Defensins 4

Abstract

牛β防御素4 (BNBD4)是一种天然存在于牛肺脏肺泡巨噬细胞内的抗菌肽，本研究拟通过克隆BNBD4成熟肽片段的编码区序列，连接至真核表达载体pPIC9K构建重组质粒mBNBD4-pPIC9K，转化至毕赤酵母菌株GS115诱导表达，镍柱纯化后，经Tricine-SDS-PAGE检测和Western Blot鉴定，成功获取重组蛋白mBNBD4，为进一步研究牛防御素BNBD4的作用机制提供试验材料。 Bovine neutrophil β-defensins 4 (BNBD4) is one of the antimicrobial peptides which naturally exists in bovine pulmonary alveolar macrophages. In this study we will clone the CDs sequence of mature BNBD4 gene, and connect it to the eukaryotic expression vector pPIC9K to construct recombinant plasmid mBNBD4-pPIC9K, and purify it by Ni-sepharose after inducing expression. Finally, we can successfully obtain the recombinant protein mBNBD4 by Tricine SDS-PAGE detection and Western Blot identify, therefore to provide the experimental materials for the further study of mature BNBD4.