Selection of cDNAs for phosphodiesterases that hydrolyze guanosine 3';5'-monophosphate in Escherichia coli.
Second Messengers Phosphoproteins
; 14(3): 127-37, 1992.
Article
em En
| MEDLINE
| ID: mdl-1345338
ABSTRACT
A genetic selection procedure has been developed which makes the growth of E. coli dependent on expression of a cGMP phosphodiesterase cDNA. E. coli does not contain a cGMP phosphodiesterase, and guanine auxotrophs cannot extract the guanine from cGMP. If a functional cGMP phosphodiesterase is introduced, then guaA auxotrophs will grow on cGMP as a guanine source. The method also selects GMP synthetase cDNAs, which complement the guanine auxotrophy directly. Expression of a Dictyostelium discoideum or human heart cyclic nucleotide phosphodiesterase cDNA permits growth of the E. coli guaA auxotroph in the presence of cGMP. Several commercial cDNA libraries were corrupt and contained phosphodiesterase and/or GMP synthetase sequences that were from a contaminating DNA source.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
DNA
/
Regulação Enzimológica da Expressão Gênica
/
3',5'-GMP Cíclico Fosfodiesterases
/
Guanosina Monofosfato
/
Escherichia coli
Limite:
Animals
/
Humans
Idioma:
En
Revista:
Second Messengers Phosphoproteins
Assunto da revista:
BIOQUIMICA
Ano de publicação:
1992
Tipo de documento:
Article