Open AccessGenetic transformation of N eisseria gonorrhoeae shows a strand preference
Author(s) -
Duffin Paul M.,
Seifert H. Steven
Publication year - 2012
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2012.02612.x
Subject(s) - neisseria gonorrhoeae , transformation (genetics) , dna , recombinant dna , palindrome , oligonucleotide , palindromic sequence , biology , microbiology and biotechnology , genetics , chemistry , gene , genome
Natural transformation is the main means of horizontal genetic exchange in the obligate human pathogen N eisseria gonorrhoeae . N eisseria spp. have been shown to preferentially take up and transform their own DNA by recognizing a non‐palindromic 10 or 12 nucleotide DNA uptake sequence ( DUS 10 or DUS 12). We investigated the ability of the DUS 12 to enhance single‐stranded DNA (ss DNA ) transformation. Given the non‐palindromic nature of the DUS 12, we tested whether both strands of the DUS equally enhance transformation. Recombinant single‐stranded M 13 phage harboring transforming DNA with the W atson DUS 12, the C rick DUS 12, or no DUS ( DUS 0) were constructed and circular ss DNA was purified. Southern blots of the purified DNA probed with strand‐specific oligonucleotide probes showed > 10 000 : 1 ratio of ss DNA to contaminating ds DNA . The C rick strand of the DUS 12 enhanced ss DNA transformation 180‐ to 470‐fold over DUS 0 ss DNA , whereas the W atson strand of the DUS only modestly enhanced ss DNA transformation in two strains of N . gonorrhoeae . These data confirm that ss DNA efficiently transforms N . gonorrhoeae , but that there is a strand preference and that part of this strand preference is a greater efficiency of the C rick strand of the DUS 12 in enhancing transformation.