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Titolo:
CONSTRUCTION OF HERMES SHUTTLE VECTORS - A VERSATILE SYSTEM USEFUL FOR GENETIC COMPLEMENTATION OF TRANSFORMABLE AND NON-TRANSFORMABLE NEISSERIA MUTANTS
Autore:
KUPSCH EM; AUBEL D; GIBBS CP; KAHRS AF; RUDEL T; MEYER TF;
Indirizzi:
MAX PLANCK INST BIOL,INFEKT BIOL ABT,SPEMANNSTR 34 D-72076 TUBINGEN GERMANY MAX PLANCK INST BIOL,INFEKT BIOL ABT D-72076 TUBINGEN GERMANY
Titolo Testata:
MGG. Molecular & general genetics
fascicolo: 5, volume: 250, anno: 1996,
pagine: 558 - 569
SICI:
0026-8925(1996)250:5<558:COHSV->2.0.ZU;2-S
Fonte:
ISI
Lingua:
ENG
Soggetto:
ESCHERICHIA-COLI; BACILLUS-SUBTILIS; EPITHELIAL-CELLS; PLASMID DNA; GONORRHOEAE; MUTAGENESIS; CLONING; TETRACYCLINE; RESISTANCE; PROTEINS;
Keywords:
PLASMID VECTOR; CONJUGATION; GENERALIZED MUTAGENESIS; HOMOLOGOUS RECOMBINATION; NATURAL TRANSFORMATION COMPETENCE;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Science Citation Index Expanded
Citazioni:
43
Recensione:
Indirizzi per estratti:
Citazione:
E.M. Kupsch et al., "CONSTRUCTION OF HERMES SHUTTLE VECTORS - A VERSATILE SYSTEM USEFUL FOR GENETIC COMPLEMENTATION OF TRANSFORMABLE AND NON-TRANSFORMABLE NEISSERIA MUTANTS", MGG. Molecular & general genetics, 250(5), 1996, pp. 558-569

Abstract

A versatile shuttle system has been developed for genetic complementation with cloned genes of transformable and non-transformable Neisseria mutants. By random insertion of a selectable marker into the conjugative Neisseria plasmid ptetM25.2, a site within this plasmid was identified that is compatible with plasmid replication and with conjugativetransfer of plasmid. Regions flanking the permissive insertion site of ptetM25.2 were cloned in Escherichia coli and served as a basis for the construction of the Hermes vectors. Hermes vectors are composed ofan E. coli replicon that does not support autonomous replication in Neisseria, e.g. ColE1, p15A, or ori(fd), fused with a shuttle consisting of a selectable marker and a multiple cloning site flanked by the integration region of ptetM25.2. Complementation of a non-transformable Neisseria strain involves a three-step process: (i) insertion of the desired gene into a Hermes vector; (ii) transformation of Hermes into aNeisseria strain containing pretM25.2 to create a hybrid ptetM25.2 via gene replacement by the Hermes shuttle cassette; and (iii) conjugative transfer of the hybrid ptetM25.2 into the final Neisseria ia recipient. Several applications for the genetic manipulation of pathogenic Neisseriae are described.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 30/10/20 alle ore 23:29:42