Cloning of PCR synthesized 191 bp DNA overlapping lac promoter for plasmid construction.
Indian J Exp Biol
; 1997 Jan; 35(1): 99-102
Article
em En
| 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:
1
Índice:
IMSEAR
Assunto principal:
Plasmídeos
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Proteínas Repressoras
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Proteínas de Bactérias
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DNA Recombinante
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Dados de Sequência Molecular
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Sequência de Bases
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Reação em Cadeia da Polimerase
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Regiões Promotoras Genéticas
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Clonagem Molecular
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Proteínas de Escherichia coli
Idioma:
En
Revista:
Indian J Exp Biol
Ano de publicação:
1997
Tipo de documento:
Article