A new vector for insertion of any DNA fragment into the chromosome of transformable Neisseriae.
A useful method for inserting any DNA fragment into the chromosome of Neisseriae has been developed. The method relies on recombination-proficient vector plasmid pNLE1, a pUC19 derivative containing (1) genes conferring resistance to ampicillin and erythromycin, as selectable markers; (2) a chromosomal region necessary for its integration into the Neisseria chromosome; (3) a specific uptake sequence which is required for natural transformation; (4) a promoter capable of functioning in Neisseria; and (5) several unique restriction sites useful for cloning. pNLE1 integrates into the leuS region of the neisserial chromosome at high frequencies by transformation-mediated recombination. The usefulness of this vector has been demonstrated by cloning the tetracycline-resistance gene (tet) and subsequently inserting the tet gene into the meningococcal chromosome.
Salvatore
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P
Dipartimento di Biologia e Patologia Cellulare e Molecolare "L. Califano,", Università di Napoli "Federico II,", Centro di Endocrinologia ed Oncologia Sperimentale "G. Salvatore" of the Consiglio Nazionale delle Ricerche, Naples, Italy.
Cantalupo
G
G
Pagliarulo
C
C
Tredici
M
M
Lavitola
A
A
Bucci
C
C
Bruni
C B
CB
Alifano
P
P
D000667:Ampicillin / Q000494:pharmacology; D002874:Chromosome Mapping; D002876:Chromosomes, Bacterial / Q000235:genetics*; D004251:DNA Transposable Elements / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics*; D004917:Erythromycin / Q000494:pharmacology; D005822:Genetic Vectors; D009345:Neisseria meningitidis / Q000235:genetics*; D010406:Penicillins / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011995:Recombination, Genetic; D013752:Tetracycline / Q000494:pharmacology; D014170:Transformation, Genetic
D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D010406:Penicillins; D011500:Protein Synthesis Inhibitors; D004917:Erythromycin; D000667:Ampicillin; D013752:Tetracycline