动物营养学报2025，Vol.37Issue(3)：2081-2089,9.DOI:10.12418/CJAN2025.176
噬菌体VB_SauM_Lm44穿孔素HolLm44的克隆表达及其抑菌效果研究
Study on Cloning and Expression of Holin HolLm44 of Bacteriophage VB_SauM_Lm44 and Its Antibacterial Effect
摘要
Abstract
This experiment aimed to clone and express of holin of Staphylococcus aureus phage VB_SauM_Lm44 and to study its antibacterial effect.Firstly,the holin gene sequence was obtained by analyzing the VB_SauM_Lm44 genome,and the holin gene(named HolLm44,the expressed perforin protein was named HolLm44)was amplified by PCR.Then,the target gene fragment was ligated with the linear pET-28a(+)plasmid after double enzyme digestion and transformed into the Escherichia coli DH5α cloned competent cells;the recombinant plasmid was extracted and then transformed into the expression strains BL21(DE3)and BL21(DE3)plysS of Escherichia coli to enable the induced expression and subsequent purification of the target pro-tein.The antibacterial effect of holin HolLm44 was evaluated through both the plate inhibition assay and the resazurin staining technique,thereby establishing the optimal conditions under which holin HolLm44 exerts its activity.The results showed that holin HolLm44 had two transmembrane regions,and its N-terminal and C-ter-minal were distributed in the cytoplasm,which belonged to classⅡ holin.The optimal temperature and pH of holin HolLm44 were 37℃and 5.2,respectively.Of the 20 Staphylococcus aureus strains used as indicator bacteria,only one was sensitive to the holin HolLm44.The results demonstrate that this study successfully cloned and expressed the active holin HolLm44 from phage VB_SauM_Lm44,confirming its potent inhibitory effect against Staphylococcus aureus.[Chinese Journal of Animal Nutrition,2025,37(3):2081-2089]关键词
噬菌体VB_SauM_Lm44/穿孔素/克隆表达/抑菌/金黄色葡萄球菌Key words
bacteriophage VB_SauM_Lm44/holin/clonal expression/antibacterial/Staphylococcus aureus分类
生物科学引用本文复制引用
张欣然,廖毅婷,熊东炜,李鹏,杨淑华,龙淼..噬菌体VB_SauM_Lm44穿孔素HolLm44的克隆表达及其抑菌效果研究[J].动物营养学报,2025,37(3):2081-2089,9.基金项目
辽宁省教育厅基本科研项目(JYTYB2024027) （JYTYB2024027）
