全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

内蒙古羊源细粒棘球蚴Eg95基因原核表达及蛋白鉴定

DOI: 10.7668/hbnxb.2010.04.012, PP. 57-60

Keywords: 细粒棘球蚴,Eg95,原核表达,蛋白纯化,生物活性

Full-Text   Cite this paper   Add to My Lib

Abstract:

在克隆内蒙古羊源细粒棘球蚴疫苗候选基因Eg95的基础上,构建原核表达载体pET-Eg95并转化E.coliBL21(DE3),优化表达体系,获得Eg95纯化蛋白.将Eg95基因从克隆质粒pMD19-T-Eg95亚克隆到原核表达载体pETa(+)上,构建原核表达载体pET-Eg95,转化E.coliBL21(DE3)诱导表达,优化表达条件.纯化的Eg95融合蛋白经SDS-PAGE和WesternBlot鉴定其正确性和免疫学活性.原核表达载体pET-Eg95在大肠杆菌中高效表达,优化的原核表达条件为:当菌液OD600值为0.6时,加入终浓度为0.1mmol/L的IPTG,37℃,振荡培养5h.纯化后的蛋白经SDS-PAGE和Westernblot鉴定为目的蛋白并具有生物学活性.原核表达载体pET-Eg95构建正确并可高效表达可溶性Nus-Eg95融合蛋白,可作为特异性抗原应用于免疫印迹法检测血清抗体.

References

[1]  蒋次鹏.我国包虫病流行近况[J].地方病通报,2002,17(3):77-79.
[2]  Lightowlers M V,Lawerence S B,Gauci C G,et al.Vaccination against hydatidosis using a defined recombinant antigen[J].Parasite Immunol,1996,18(9):457-462.
[3]  Lightowlers M V,Jensen O,Fernandez E,et al.Vaccination trials in Australia and Argentina confirm the effectiveness of the Eg95 hydatid vaccine in sheep[J].Int J Parasitol,1999,29:531-534.
[4]  Chow C,Charles G G,Alan F C,et al.A gene family expressing a host-protective antigen of Echinococcus granulosus[J].Mol Biochem Parasitol,2001,118(1):83-88.
[5]  潘卫庆,汤林华.分子寄生虫学[M].上海:上海科学技术出版社,2004:345-399.
[6]  李志伟,王志钢.细粒棘球蚴内蒙株疫苗候选基因 Eg95 克隆[J].中国公共卫生,2008,24(1):56-57.
[7]  Jenkins R E,Tanner J A.The structure of the major protein of the human erythrocyte membrane.Characterization of the intact protein and major fragments[J] Biochem J,1977,161(1):139-147.
[8]  Deplazes P,Jimenez-Palacios S,Gottstein B,et al.Detection of Echinococcus coproantigens in stray dogs of northern Spain[J].Appl Parasitol,1994,35(4):297-301.
[9]  Deplazes P,Thompson R C,Constantine C C,et al.Primary infection of dogs with Echinococcus granulosus:systemic and local (Peyer′s patches) immune responses[J].Vet Immunol Immunopathol,1994,40(2):171-184.
[10]  Rigano P,Manfre L,La Galla R,et al.Clinical and hematological response to hydroxyurea in a patient with Hb Lepore/beta-thalassemia[J].Hemoglobin,1997,21(3):219-26.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133