Antimicrobial Effect of Zophobas morio defensin against Bovine Mastitis Pathogens
作者单位：College of Veterinary Medicine China Agricultural University
会议名称：《中国畜牧兽医学会兽医产科学分会第八届会员代表大会暨第十五次学术研讨会》
会议日期：2021年
学科分类：090603[农学-临床兽医学] 09[农学] 0906[农学-兽医学]
摘 要：Objective:Mastitis has always been considered as a great threat for dairy cow industry around the ***,mastitis treatment mainly relies on ***,many countries restrict the use of antibiotics since the bacterial resistance and drug residues have become a serious issue in the past decades of ***,it is urgent to develop a broad-spectrum,low residual anti-bacterial drug against bacterial resistance and drug ***:We analyzed the active ingredient of Zophobas morio(***) hemolymph and obtained the full-length complete coding sequence(CDs) of *** defensin through 5’ and 3’ RACE *** target gene was cloned into pGEX 6 p-1 and transformed into *** Rosetta(DE3) *** protein expression was induced with isopropyl-p-D-1-thiogalactopyranoside(IPTG),and purified through GSTrap 4 ***,the recombinant protein was treated with Prescission Protease and purified by Superdex 30 to remove glutathione S-transferase(GST)-***-zone assay was carried out to evaluate the antimicrobial activity of recombinant *** assay and cytotoxicity test were used to evaluate the safety of recombinant ***:Our results showed that the gene of Z. morio defensin encodes 89 amino acids,and the structure of *** defensin protein includes a conformationally flexible amino-terminal loop(n-loop) followed by a cysteinestabilized α-helical and β-sheet(CSαβ) domain,which is a structural scaffold shared by defensins of other *** addition,recombinant Z. morio defensin protein could inhibit multiple bacteria growths in vitro,such as Escherichia coli,Staphylococcus aureus,Klebsiella pneumonia,bacillus cereus,et ***,recombinant Z. morio defensin protein also maintains non-hemolytic and non-cytotoxicity as ***:In summary,recombinant Z. morio defensin protein which showed excellent antibacterial activity and safety,is a promising drug candidate to treat mastitis for dairy cows.