Gene cloning and expression and characterization of a toxin-sensitive protein phosphatase from the methanogenic archaeon Methanosarcina thermophila TM-1.
نویسندگان
چکیده
With oligonucleotides modelled after conserved regions within the protein-serine/threonine phosphatases (PPs) of the PP1/2A/2B superfamily, the gene for the archaeal protein phosphatase PP1-arch2 was identified, cloned, and sequenced from the methanogenic archaeon Methanosarcina thermophila TM-1. The DNA-derived amino acid sequence of PP1-arch2 exhibited a high degree of sequence identity, 27 to 31%, with members of the PP1/2A/2B superfamily such as PP1-arch1 from Sulfolobus solfataricus, PP1alpha from rats, PP2A from Saccharomyces cerevisiae, and PP2B from humans. The activity of the recombinant PP1-arch2 was sensitive to several naturally occurring microbial toxins known to potently inhibit eucaryal PP1 and PP2A, including microcystin-LR, okadaic acid, tautomycin, and calyculin A.
منابع مشابه
The methanogenic archaeon Methanosarcina thermophila TM-1 possesses a high-affinity glycine betaine transporter involved in osmotic adaptation.
Methanogenic Archaea are found in a wide range of environments and use several strategies to adjust to changes in extracellular solute concentrations. One methanogenic archaeon, Methanosarcina thermophila TM-1, can adapt to various osmotic conditions by synthesis of alpha-glutamate and a newly discovered compatible solute, Ne-acetyl-beta-lysine, or by accumulation of glycine betaine (betaine) a...
متن کاملPCR-MEDIATED CLONING A ND EXPRESSION OF THE GENE FOR THE B-SUBUNIT OF VIBRIO CHOLERAE TOXIN ISOLATED RECENTLY IN IRAN
Knowing the nucleotide sequence of the cholera toxin operon, we designed oligonucleotide primers for its-PCR amplification from local clinical isolates of V. cholerae. The resulting amplification product was cloned in a common pUC18 vector. Subsequently, a part of this operon encoding the cholera toxin Bsubunit (CTB) was reamplified and cloned between the BamH1 and EcoR1 sites of the same ...
متن کاملIn silico fusion of epsilon and beta toxin genes of Clostridium perfringens types D and B
Fusion protein technology represents the strategy to achieve rapid, efficient, and cost-effective proteinexpression. Epsilon and Beta toxins are the most potent Clostridial toxins and cause disease in animals.This study describes in silico fusion of Clostridium perfringens types D and B epsilon and beta toxin genesthat was used for cloning in E.coli. The etx and cpb genes were...
متن کاملFusion 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...
متن کاملCloning and characterization of MAP2191 gene, a mammalian cell entry antigen of Mycobacterium avium subspecies paratuberculosis
The aim of this study is to identify, clone and express a Mycobacterium avium subsp. paratuberculosis specific immunogenic antigen candidate, in order to develop better reagents for diagnosis and vaccines for the protection of the host. Therefore, MAP2191 gene (a member of MAPmce5 operon) from MAP, was isolated and characterized by Bioinformatics tools and <e...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- Journal of bacteriology
دوره 179 16 شماره
صفحات -
تاریخ انتشار 1997