Open AccessGenetic Manipulation of Corynebacterium diphtheriae and Other Corynebacterium Species
Author(s) -
Chang Chungyu,
Nguyen Minh Tan,
TonThat Hung
Publication year - 2020
Publication title -
current protocols in microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.813
H-Index - 35
eISSN - 1934-8533
pISSN - 1934-8525
DOI - 10.1002/cpmc.111
Subject(s) - corynebacterium diphtheriae , biology , corynebacterium glutamicum , corynebacterium , transposable element , mutagenesis , transposon mutagenesis , genetics , complementation , mutant , levansucrase , shuttle vector , gene , diphtheria , bacteria , virology , bacillus subtilis , vaccination , vector (molecular biology) , recombinant dna
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
Accelerating Research
Robert Robinson Avenue,
Oxford Science Park, Oxford
OX4 4GP, United Kingdom
Address
John Eccles HouseRobert Robinson Avenue,
Oxford Science Park, Oxford
OX4 4GP, United Kingdom