The Morganella morganii phoC gene, encoding a class A acid phosphatase, was used to generate gene fusions with modified amino-terminal moieties of the Escherichia coli lacZ gene carrying a multiple-cloning site flanked by phage-specific promoters and recognition sites for universal sequencing primers. The corresponding hybrid proteins retained a PhoC-like enzymatic activity which is easily detectable by a plate histochemical assay, rendering similar gene fusions potentially useful as targets for cloning-dependent insertional inactivation. Cloning experiments performed in plasmids carrying similar lacZ-phoC fusions confirmed their usefulness as cloning vectors for direct screening of recombinants. As compared to conventional lacZ alpha-complementation-based vectors, which can only be used in E. coli hosts carrying specific lacZ mutations, the lacZ-phoC fusion-based vectors can be used in combination with any E. coli host and require a less expensive histochemical assay for screening of recombinants, while retaining all the advantageous features that made the former so popular as general purpose cloning vehicles.

Thaller, M.c., Berlutti, F., Schippa, S., Selan, L., Rossolini, G. (1998). Bacterial acid phosphatase gene fusions useful as targets for cloning-dependent insertional inactivation. BIOTECHNOLOGY PROGRESS, 14(2), 241-247 [10.1021/bp980009t].

Bacterial acid phosphatase gene fusions useful as targets for cloning-dependent insertional inactivation

THALLER, MARIA CRISTINA;
1998-01-01

Abstract

The Morganella morganii phoC gene, encoding a class A acid phosphatase, was used to generate gene fusions with modified amino-terminal moieties of the Escherichia coli lacZ gene carrying a multiple-cloning site flanked by phage-specific promoters and recognition sites for universal sequencing primers. The corresponding hybrid proteins retained a PhoC-like enzymatic activity which is easily detectable by a plate histochemical assay, rendering similar gene fusions potentially useful as targets for cloning-dependent insertional inactivation. Cloning experiments performed in plasmids carrying similar lacZ-phoC fusions confirmed their usefulness as cloning vectors for direct screening of recombinants. As compared to conventional lacZ alpha-complementation-based vectors, which can only be used in E. coli hosts carrying specific lacZ mutations, the lacZ-phoC fusion-based vectors can be used in combination with any E. coli host and require a less expensive histochemical assay for screening of recombinants, while retaining all the advantageous features that made the former so popular as general purpose cloning vehicles.
1998
Pubblicato
Rilevanza internazionale
Articolo
Esperti anonimi
Settore BIO/19 - MICROBIOLOGIA GENERALE
Settore MED/07 - MICROBIOLOGIA E MICROBIOLOGIA CLINICA
English
Con Impact Factor ISI
Acid Phosphatase; Artificial Gene Fusion; Cloning, Molecular; DNA, Recombinant; Escherichia coli; Genetic Code; Genetic Vectors; Lac Operon; Plasmids; Genes, Bacterial; Mutagenesis, Insertional
Thaller, M.c., Berlutti, F., Schippa, S., Selan, L., Rossolini, G. (1998). Bacterial acid phosphatase gene fusions useful as targets for cloning-dependent insertional inactivation. BIOTECHNOLOGY PROGRESS, 14(2), 241-247 [10.1021/bp980009t].
Thaller, Mc; Berlutti, F; Schippa, S; Selan, L; Rossolini, G
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2108/175520
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