title: [Construction of Staphylococcus aureus RN6390 hmp gene mutant and analysis of NO sensitivity].
abstract: OBJECTIVE
  To investigate the function of flavohaemoglobin (HMP) in Staphylococcus aureus RN6390 under the nitrification pressure, we constructed the hmp gene deletion mutant of RN6390 strain.
  
  
  METHODS
  According to principle of homologous recombination, we obtained the up stream and down stream sequences of hmp gene by PCR using chromosomal DNA of S. aureus RN6390 as template. Antibiotics pressure and alternating temperature culture were applied for mutant strain selection. We verified the clones screened out by genome PCR and real-time PCR quantification. Sodium nitroprusside (SNP), as nitric oxide (NO) donor, was used for NO resistance evaluation. In addition, we compared the bacteria biofilm formation ability of hmp gene mutant strain with wild type.
  
  
  RESULTS
  We successfully constructed hmp gene mutant strain of S. aureus RN6390. The expression of hmp gene was direct correlate with the concentration of exogenous NO. We found that compared to wild type, the mutant strain was more sensitive to NO and it is prone to form bacteria biofilm.
  
  
  CONCLUSIONS
  The successfully constructed hmp gene deletion mutant of S. aureus provided the possibilities to further investigate the biological function of hmp gene in the resistance of S. aureus to NO from host immune system.
authors.affiliation: Key Laboratory of Marine Drugs, Chinese Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, China. jp886@163.com
authors.forename: Peng
authors.initials: P
authors.lastname: Jiang
authors.forename: Xinzhi
authors.initials: X
authors.lastname: Lu
authors.forename: Fangjie
authors.initials: F
authors.lastname: Hou
authors.forename: Zhanjun
authors.initials: Z
authors.lastname: Lin
authors.forename: Wengong
authors.initials: W
authors.lastname: Yu
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D006420:Hemeproteins / Q000235:genetics* / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism*; D017384:Sequence Deletion; D013211:Staphylococcus aureus / Q000235:genetics* / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D006420:Hemeproteins; C086193:flavohemoprotein, Bacteria; D009569:Nitric Oxide