【摘 要】
：
目的 克隆、表达脑多头蚴超氧化物歧化酶(Cu/Zn-SOD)基因并分析其酶学活性.方法 从羊脑多头蚴原头节提取总RNA,采用RT-PCR技术扩增脑多头蚴超氧化物歧化酶(Cu/Zn-SOD)基因,将其克隆入原核表达载体pET-32a(+)中,构建pET-32a-Cu/Zn-SOD表达载体,转化大肠杆菌BL21(DE3)后诱导表达,用SDS-PAGE和Western-blot检测表达产物,并对Cu/Z
【机 构】
：
四川农业大学动物医学院动物寄生虫病研究中心,成都 611130;成都农业科技职业学院,成都 611130
【出 处】
：
中国畜牧兽医学会兽医寄生虫学分会第十三次学术研讨会
论文部分内容阅读
目的 克隆、表达脑多头蚴超氧化物歧化酶(Cu/Zn-SOD)基因并分析其酶学活性.方法 从羊脑多头蚴原头节提取总RNA,采用RT-PCR技术扩增脑多头蚴超氧化物歧化酶(Cu/Zn-SOD)基因,将其克隆入原核表达载体pET-32a(+)中,构建pET-32a-Cu/Zn-SOD表达载体,转化大肠杆菌BL21(DE3)后诱导表达,用SDS-PAGE和Western-blot检测表达产物,并对Cu/Zn-SOD重组蛋白进行酶学活性分析.结果 结果显示Cu/Zn-SOD基因ORF框为459bp,编码152个氨基酸,推导的蛋白分子大小为16.72kDa,与亚洲带绦虫、猪带绦虫、肥头绦虫Cu/Zn-SOD的基因序列相似性分别为99％、98％和92％.纯化的原核表达产物中Cu/Zn-SOD蛋白的含量为1.587mg.mL-1,酶的比活力为3138IU/mg.结论本研究成功表达了脑多头蚴Cu/Zn-SOD基因并鉴定了它的酶学活性,为进一步研究Cu/Zn-SOD在脑多头蚴中的生物学功能奠定了基础.
其他文献
(Objective)This experiment was conducted to investigate Trichomonas spp.infection in pigeons of Shandong Province.(Result)Oropharyngeal swabs(n =609)were collected randomly from 80,000 domestic pigeon
Cancer is a general name for more than 100 malignant diseases.It is postulated that all cancers start from a single abnormal cell that grows out of control.Untreated cancers can cause serious conseque
Background: Hco-gal-m and-f were two isoforms of galectin cloned from male and female Haemonchus contortus,respectively,and it was demonstrated that recombinant Hco-gal-m and-f could act as immune sup
In this study,a combined proteomic and transcriptomic analysis was performed to understand the mechanisms underlying the immunomodulation induced by recombinant galectins of Haemonchus contortus(rHco-
The specific primers derived from T.spiralis serine proteinase inhibitor(TsSerPIN)gene in GenBank database were designed and used to amplify the TsSerPIN gene from total RNA isolated from T.spiralis m
Trichinella spiralis,an intracellular parasite nematode,can cause severe foodborne zoonosis that is known as tichinellosis.The life cycle of T.spiralis consist of adult(Ad),muscle larvae(ML)and newbor
Trichinellosis is a an emerging and reemerging infectious disease,caused by Trichinella spp.Cathepsin F belongs to cysteine protease that is a major virulence factor for parasitic helminths,and it may
Nematophagous fungi are considered to have the best potential as biological agents for the control of gastrointestinal nematodes in domestic animals.However,relatively few studies have been conducted
Vaccination as one of prevention strategies against the Trichinella spiralis infection in hosts is an ongoing effort.However,no ideal vaccine candidates have been identified until now.Therefore,identi
目的 克隆和原核表达脑多头蚴dUTPase基因并分析其酶学活性.方法 从羊脑多头蚴原头节提取总RNA,采用RT-PCR技术扩增脑多头蚴dUTPase基因,将其克隆入原核表达载体pET-32a(+)中,构建pET-32a-dUTPase表达载体,转化大肠杆菌BL21(DE3)后诱导表达,用SDS-PAGE和Western-blot检测表达产物,用免疫荧光组化法检测dUTPase基因在多头带绦虫成虫体