Insects protect themselves against microbial infection by an efficient innate immune system that is activated by recognition ofinvariant microbial surface molecules. In the fruit fly Drosophila melanogaster the presen...Insects protect themselves against microbial infection by an efficient innate immune system that is activated by recognition ofinvariant microbial surface molecules. In the fruit fly Drosophila melanogaster the presence of bacteria is associated with expression of antimicrobial peptides in host immune-competent tissues. Host receptors detect infection and relay the signal to mount the appropriate immune response. In Drosophila hemocyte- like l（2）mbn cells pre-infection treatment with Pefabloc, a commonly used serine protease inhibitor, induced two major effects： it blocked expression of the antibacterial peptide Diptericin in response to live Gram-negative bacteria and bacterial surface molecules （crude lipopolysaccharide contaminated by peptidoglycans） and it induced morphological changes.展开更多
文摘为了实现原核表达的途径高效获取抗菌肽蛋白,以RT-PCR的方法反转录合成家蝇的抗菌肽diptericin基因,并克隆至p GEX-4T-1载体上,转化至大肠杆菌BL21宿主菌进行表达。测序结果显示,RT-PCR克隆到长345 bp的家蝇diptericin基因,重组菌经异丙基-β-D-硫代半乳糖苷(IPTG)诱导后,通过SDS-PAGE电泳和Western-blot检测到目的蛋白的融合表达,结果表明带有GST标签的融合蛋白大小约为37 k D,并且通过GST纯化柱纯化得到了目的蛋白。
文摘Insects protect themselves against microbial infection by an efficient innate immune system that is activated by recognition ofinvariant microbial surface molecules. In the fruit fly Drosophila melanogaster the presence of bacteria is associated with expression of antimicrobial peptides in host immune-competent tissues. Host receptors detect infection and relay the signal to mount the appropriate immune response. In Drosophila hemocyte- like l（2）mbn cells pre-infection treatment with Pefabloc, a commonly used serine protease inhibitor, induced two major effects： it blocked expression of the antibacterial peptide Diptericin in response to live Gram-negative bacteria and bacterial surface molecules （crude lipopolysaccharide contaminated by peptidoglycans） and it induced morphological changes.
