黑腹果蝇抗真菌肽基因Drs和Drs-lC在原核中的表达及抗菌活性的初步测定  

Expression of the Antifungal Peptide Genes,Drs and Drs-lC From Drosophila melanogaster in E.coli and the Activity Detection of Their Expression Products

作　　者：邓小娟[1] 桑延霞[1] 杨婉莹[1] 钟仰进[1] 黄亚东[2] 曹阳[1] 温硕洋[3] 

机构地区：[1]华南农业大学动物科学学院蚕丝科学系,广州510642 [2]暨南大学医药生物技术研究开发中心,广州510632 [3]华南农业大学资源与环境学院昆虫学系,广州510642

出　　处：《蚕业科学》2006年第2期264-267,共4页ACTA SERICOLOGICA SINICA

基　　金：国家重点基础研究发展计划"973"项目(编号2005CB121000);广东省科学技术计划重点项目(编号2003C104042);广东省自然科学基金项目(编号010290;032256;04020553)

摘　　要：对黑腹果蝇(Drosophila m elanogaster)抗真菌肽基因Drs和它的同系物基因Drs-lC在pET-21d载体中与T7.Tag标签序列进行了融合表达,并对融合表达产物进行Xa因子切割前后的抗菌活性测定。首先通过长引物PCR获得5′端带有Xa因子识别切割序列的Drs和Drs-lC,分别克隆至pET-21d表达载体上,然后转化E.coliO rigam i(DE3),筛选得到阳性克隆。以IPTG诱导目的基因与载体上的T7.Tag标签序列融合表达,将表达产物进行初步纯化后,以Xa因子进行切割处理,然后测定处理前后样品的抗菌活性。结果显示与T7.Tag标签序列融合表达的D rs、D rs-lC样品和经Xa因子切割后的样品对供试真菌黄色镰刀菌(Fusarium culm orum)、尖孢镰孢菌(Fusarium oxs-porum)和粗糙脉孢菌(Neuropora crassa)均有明显的抑制作用。The antifungal peptide genes, Drs and Drs-IC from Drosophila melanogaster were cloned into the pET-21d vector and fusion expressed with T7·Tag in E. coll. The antifungal activity of expression product was detected. Firstly, the target genes, Drs and Drs-lC with a cleavage site of Xa factor at 5＇ end were obtained by PCR with long primers, and then were cloned into the pET-21d vector. The recombinant vectors, pET-21d-xa-Drs and pET-21d-xa-lC were transformed into the host cells of E. coli Origami （DE3）. Drs, Drs-IC fused with T7·Tag were induced to express by IPTG, The expression products were purified and digested by Xa factor, The result showed that the fore- and-aft samples by digestion of Xa factor had obvious antifungal activity to the tested fungi, Fusarium culmorum, Fusarium oxsporum and Neuropora crassa.

关 键 词：黑腹果蝇 抗真菌肽基因 DRS Drs-lC 融合表达 抗菌活性 

分 类 号：Q966[生物学—昆虫学] Q78