cloning and expression of s1 subunit of pertussis toxin in escherichia coli
Authors
abstract
bordetella pertussis is a gram negative bacterium that causes respiratory tract infection in human (whooping cough). pertussis toxin (pt) is the main component of current acellular pertussis vaccine and the s1 (subunit1) is the main immunogenic part of it. thus, s1 has been the target of many studies as a potent candidate of acellular vaccine against bordetella pertussis, lacking the side effects of whole cell based ones. s1 gene was amplified and inserted in three expression vectors including pet-14, pet-22b(+) and paed4. the possibility and level of expression of these constructs were investigated in bl21 (de3) strain of escherichia coli (e.coli) as expression host. the highest expression was in pet-22b(+)-s1. best expression achieved 6 hr post induction with 0.2 mm iptg in lb broth containing ampicillin, at 30°c with shaking (250 rpm). recombinant s1 protein was observed in two distinct separated proteins with 28 and 31 kda estimated molecular weight. in spite of toxicity of pt and s1 in the e.coli, considerable amount of s1 was expressed in e.coli. two rs1 bands were detected on sds-page. both were confirmed as s1 in western blot with specific monoclonal and polyclonal antibodies against pertussis toxin. appearance of two distinct bands could be the result of leader peptidase activity or nonspecific peptidase from e.coli on recombinant s1. as the recombinant s1 is a suitable antigen for studies as a candidate acellular vaccine or development of elisa for detection of bordetella pertussis, further studies are underway.
similar resources
Cloning and Expression of S1 Subunit of Pertussis Toxin in Escherichia coli
Bordetella pertussis is a gram negative bacterium that causes respiratory tract infection in human (whooping cough). Pertussis toxin (PT) is the main component of current acellular pertussis vaccine and the S1 (subunit1) is the main immunogenic part of it. Thus, S1 has been the target of many studies as a potent candidate of acellular vaccine against Bordetella pertussis, lacking the side effec...
full textCloning of Clostridium perfringens iota toxin gene in Escherichia coli
Iota toxin is produced by Clostridium perfringens type E. This toxin causes antibiotic-associated enterotoxemia in lambs and calves. Iota toxin is a binary toxin that has two components including Ia (the enzyme component) and Ib (the binding component). Ib binds to the surface receptor of target cells and translocate Ia into the cytosol of cells. The aim of this study was to clone toxigenic epi...
full textFusion of Cholera toxin B subunit (ctxB) with Shigella dysenteriae type I toxin B subunit (stxB), Cloning and Expression that in E. coli
Background and Objective: Shiga toxin (STx) is the main virulence factor in Shigella Dysenteriae type I and is composed of an enzymatic subunit STxA monomer and a receptor-binding STxB homopentamer. Shigella toxin B subunit (STxB) is a non-toxic homopentameric protein responsible for toxin binding and internalization into target cells by interacting with glycolipid (Gb3). Cholera toxi...
full textCloning and evaluation of gene expression and purification of gene encoding recombinant protein containing binding subunit of coli surface antigens CS1 and CS2 from Enterotoxigenic Escherichia coli
Background & Objective: Enterotoxigenic Escherichia coli (ETEC) is a major causative agent of diarrhea. Enterotoxins and the colonization factors (CFs) are major virulence factors in ETEC infections. The bacterium binds to the intestinal epithelial cell surface through colonization factors and produces enterotoxins that cause excessive fluid and electrolyte secretion in the lumen of the intesti...
full textMembrane localization of the S1 subunit of pertussis toxin in Bordetella pertussis and implications for pertussis toxin secretion.
Pertussis toxin is secreted from Bordetella pertussis with the assistance of the Ptl transport system, a member of the type IV family of macromolecular transporters. The S1 subunit and the B oligomer combine to form the holotoxin prior to export from the bacterial cell, although the site of assembly is not known. To better understand the pathway of pertussis toxin assembly and secretion, we exa...
full textMy Resources
Save resource for easier access later
Journal title:
avicenna journal of medical biotechnologyجلد ۳، شماره ۱، صفحات ۱۹-۲۴
Keywords
Hosted on Doprax cloud platform doprax.com
copyright © 2015-2023