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Titolo:
Construction of a chimeric shuttle plasmid via a heterodimer system: Secretion of an scFv protein from Bacillus brevis cells capable of inhibiting hemagglutination
Autore:
Shiroza, T; Shibata, Y; Hayakawa, M; Shinozaki, N; Fukushima, K; Udaka, S; Abiko, Y;
Indirizzi:
Nihon Univ, Sch Dent, Dept Biochem, Matsudo, Chiba 2718587, Japan Nihon Univ Matsudo Chiba Japan 2718587 hem, Matsudo, Chiba 2718587, Japan Nihon Univ, Sch Dent, Dept Microbiol, Matsudo, Chiba 2718587, Japan Nihon Univ Matsudo Chiba Japan 2718587 iol, Matsudo, Chiba 2718587, Japan Tokyo Univ Agr, Dept Fermentat Sci, Setagaya Ku, Tokyo 1568502, Japan Tokyo Univ Agr Tokyo Japan 1568502 ci, Setagaya Ku, Tokyo 1568502, Japan
Titolo Testata:
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY
fascicolo: 2, volume: 65, anno: 2001,
pagine: 389 - 395
SICI:
0916-8451(200102)65:2<389:COACSP>2.0.ZU;2-3
Fonte:
ISI
Lingua:
ENG
Soggetto:
BIOLOGICALLY-ACTIVE FORM; HIGH-LEVEL SECRETION; ORAL STREPTOCOCCI; TRANSFORMATION; ANTIBODY; STRAIN; GENES;
Keywords:
scFv secretion; Bacillus brevis; pUB110; inhibition of hemagglutination; passive immunization;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Life Sciences
Citazioni:
17
Recensione:
Indirizzi per estratti:
Indirizzo: Shiroza, T Nihon Univ, Sch Dent, Dept Biochem, Matsudo, Chiba 2718587, Japan Nihon Univ Matsudo Chiba Japan 2718587 o, Chiba 2718587, Japan
Citazione:
T. Shiroza et al., "Construction of a chimeric shuttle plasmid via a heterodimer system: Secretion of an scFv protein from Bacillus brevis cells capable of inhibiting hemagglutination", BIOS BIOT B, 65(2), 2001, pp. 389-395

Abstract

Passive immunization is an attractive therapy for preventing oral diseasesincluding dental caries and periodontal disease. For this purpose, we attempted to produce a single chain variable fragment, scFv, which inhibited hemagglutination using the Bacillus brevis protein-producing system. To accomplish this, a novel strategy, a heterodimer system, was used for the construction of a chimeric shuttle plasmid. Initially, a set of new plasmids, kanamycin-resistant donor and erythromycin-resistant general cloning plasmids,were constructed. p15A ori was a common replication origin in these plasmids, while the pUB110 rep and minus origin (MO) were cloned into the donor plasmid. Next, the secretion domain of the B. subtilis alpha -amylase gene and the G2-4 gene, coding for the scFv protein, were cloned into the generalcloning plasmid and fused by PCR. Both the donor plasmid and the general cloning plasmid containing the fused gene were digested with Not1 and them ligated, a dimeric plasmid being constructed. The hey restriction sites, Asci, are arranged such that the pUB110 rep-MO moiety was switched from the donor to the general cloning plasmid following Asci digestion. The chimeric shuttle plasmid was readily constructed by simple re-circularization and a B. brevis transformant producing the scFv protein in the culture fluid was isolated.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 05/04/20 alle ore 00:22:12