In vitro recombination and identification of mutated fragment corresponding to regulation region of mtrR gene of Neisseria gonorrhoeae.
A site-directed mutant DNA fragment was synthesized and transfected into clinical Neisseria gonorrhoeae (NG) stains to construct the transformants that contained the corresponding mutagenesis of regulation region of mtrR gene. According to the technique of gene splicing by overlap extension (SOEing), a DNA segment with specific mutagenesis was constructed by two-step polymerase chain reaction (PCR). The mutation fragments EF could be used for the next experiment in which the mutation NG strains were induced. By comparing the recombinant EF fragments to the corresponding DNA fragments of clinical NG strains, 2 of these were not compatible completely. The results of sequencing revealed that there was a 9 bp deletion between the 45 to 54 inverted repeat sequence localized within the mtrR promoter. It can be confirmed that the fragments EF are the specifically designed mutant fragments.
Department of Dermatology, Union Hospital, Tongji Medical College of Huazhong University of Science and Technology, Wuhan, China. hcz0501@126.com
Changzheng
C
Huang
Nengxing
N
Lin
Yating
Y
Tu
Xin
X
Lian
Jian
J
Kang
Li
L
Zhu
D001426:Bacterial Proteins / Q000235:genetics*; D053938:DNA Fragmentation; D004269:DNA, Bacterial / Q000235:genetics; D016297:Mutagenesis, Site-Directed; D009344:Neisseria gonorrhoeae / Q000235:genetics* / Q000378:metabolism; D011995:Recombination, Genetic; D012097:Repressor Proteins / Q000235:genetics*; D017384:Sequence Deletion; D014162:Transfection; D014169:Transformation, Bacterial
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D012097:Repressor Proteins; C087450:mtrR protein, Neisseria gonorrhoeae