排序方式: 共有11条查询结果,搜索用时 2 毫秒
11.
Alvandi Amirhooshang Farajzadeh Ahmad Ghaforian Borojerdnia Mehri Jelodar Abbass Aryan Ehsan Gholipour Abolfazl Masjedizadeh Abdolrahim Makvandi Manoochehr 《World journal of microbiology & biotechnology》2011,27(4):969-974
UreB is one of the urease subunits of Helicobacter pylori and can be used as an excellent antigen candidate for H. pylori vaccination. Easy access to highly purified UreB protein, facilitate advances in therapeutic or preventive strategies. To
achieve a simplified purification procedure, the present report represents a novel method of producing recombinant urease
subunit B extracellularly. ureB gene from 26,695 standard strain was amplified by PCR and cloned into pET-26b(+) expression vector. UreB was expressed as
a soluble, N-terminal pelB and C-terminal hexahistidine-tagged fusion protein (UreB-6His) and secreted into the periplasmic
space of Escherichia coli. Expression of the recombinant UreB in E. coli BL21 (DE3) was induced by isopropylthio-β-d-galactoside (IPTG). Expression of UreB was confirmed by sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE)
and western blot analysis using anti-His monoclonal antibody. UreB-6His protein was extracted from the periplasm by osmotic
shock treatment and was purified in one step by Nickel affinity chromatography. In conclusion, the present protocol is easier
to perform; more time effective and low cost than earlier methods. 相似文献