Cloning of PCR synthesized 191 bp DNA overlapping lac promoter for plasmid construction.
Indian J Exp Biol
;
1997 Jan; 35(1): 99-102
Artigo
em Inglês
| IMSEAR
| ID: sea-56530
ABSTRACT
E coli genomic DNA was amplified by polymerase chain reaction (PCR) using two 5' and 3' oligonucleotide primers (27-mer). Amplified DNA was 191 bp. The region of amplified DNA on lac operon in E coli was 14 bp upstream from the transcription initiation site and 177 bp downstream. Amplified DNA was cloned into a phagemid vector for construction of plasmid, suitable for use as template for making strand-specific probe to detect initiated lac transcript by RNase protection assay, labelling for Southern and Northern hybridization. Another use of this probe made from this construct is to detect strand-specific DNA repair. The construct was verified by DNA sequencing.
Texto completo:
DisponíveL
Índice:
IMSEAR (Sudeste Asiático)
Assunto principal:
Plasmídeos
/
Proteínas Repressoras
/
Proteínas de Bactérias
/
DNA Recombinante
/
Dados de Sequência Molecular
/
Sequência de Bases
/
Reação em Cadeia da Polimerase
/
Regiões Promotoras Genéticas
/
Clonagem Molecular
/
Proteínas de Escherichia coli
Idioma:
Inglês
Revista:
Indian J Exp Biol
Ano de publicação:
1997
Tipo de documento:
Artigo
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