Genetic Manipulation of Corynebacterium diphtheriae and Other Corynebacterium Species.
Curr Protoc Microbiol
; 58(1): e111, 2020 09.
Article
em En
| MEDLINE
| ID: mdl-32865881
ABSTRACT
This article describes several established approaches for genetic manipulation of Corynebacterium diphtheriae, the causative agent of diphtheria that is known to have provided key evidence for Koch's postulates on the germ theory. First, it includes a detailed gene deletion method that generates nonpolar, in-frame, markerless deletion mutants, utilizing the levansucrase SacB as a counter-selectable marker. Second, it provides a thorough protocol for rescuing deletion mutants using Escherichia coli-Corynebacterium shuttle vectors. Finally, a Tn5 transposon mutagenesis procedure is described. In principle, these protocols can be used for other Corynebacterium species, including Corynebacterium glutamicum and Corynebacterium matruchotii. © 2020 Wiley Periodicals LLC Basic Protocol 1 Gene deletion in Corynebacterium diphtheriae Basic Protocol 2 Complementation of a mutant strain Basic Protocol 3 Tn5 transposon mutagenesis of Corynebacterium diphtheriae.
Palavras-chave
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Mutagênese Insercional
/
Corynebacterium
/
Corynebacterium diphtheriae
/
Teste de Complementação Genética
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article