Kowsar Medical Journal Vol. 16, No. 3, Fall 2011 Pages: 145-149
saCOL2291 saCOL2581
PhD
*
PhD PhD
MSc PhD *
saCOL2581 saCOL2291
CSF PCR
DNA
Gene runner
Puls4
N C C saCOL
Gene polymorphism of saCOL2291 and saCOL2581; candidate genes for vaccine in Methicillin-resistant Staphylococcus aureus Rouhani Nejad H.1 MSc, Fallah Mehrabadi J.2 PhD, Pourmand M. R.* PhD, Zolfaghari M. R.1 PhD, Imani Fooladi A. A.3 PhD ٭Department of Microbiology, Faculty of Health, Tehran University of Medical Sciences, Tehran, Iran 1 Department of Microbiology, Faculty of Basic Sciences, Qom Branch, Islamic Azad University, Qom, Iran 2 MARS Bioinformatics Institute, Tehran, Iran 3 Applied Microbiology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran
Abstract Aims: Staphylococcus aureus is an important nosocomial infectious agent. The prevalence of antibiotic resistance complicates the treatment of staphylococcal infections. Therefore, the development of an effective vaccine against Staphylococcus aureus is important. saCOL2291 and saCOL2581 are potential candidate genes for vaccine development and their sequences are available in Gene Bank. This study was carried out with the aim of determining the mutations in different isolates and confirming the protein of these genes. Materials & Methods: Genomic DNA was extracted from 30 clinical Staphylococcus aureus isolates, randomly obtained from urine, ulcer, mucosa and CSF samples of the inpatients of different Iranian hospitals. The genomic DNA was extracted after culture and the genes were PCR-amplified. The amplicons were sequenced and aligned with Puls4 software. Then, the nucleotide sequences were changed into amino-acid sequences by Gene runner software and the protein sequences were aligned and analyzed. Results: Nucleotide polymorphisms resulted in amino-acid sequence changes. These polymorphisms were located in the N-terminal domains, while the C-terminal domains of these genes were conserved. Conclusion: Both genes are appropriate candidates for vaccine due to their presence in most Staphylococcus aureus species and the existence of conserved C-terminal domain. Keywords: Vaccine, saCOL, Staphylococcus aureus
. فزستبده شوند
[email protected] تمبم درخواستهب ببید به نشبنی. محمدرضب پورمند:نویسنده مسئول 00/3/11 : پذیزش مقبله00/1/22 :دریبفت مقبله
PVL (egc)
C
Sca Sca
saCOL2581
ScaD
MRSA
ScaC
saCOL2291
mec
saCOL2291 saCOL2581
CSF DNase ABC DNA LB DNA
SA2006 FnbpB
FnbpA
A
SA1472 SA2291
saCOL2291 DNA
saCOL2581 DNA Pfu
A
(NC_2951.2) PCR (NC_002951.2) saCOL2581 saCOL2291 NCBI PCR
COL
TSST-1
saCOL2291 saCOL2581
saCOL2581 saCOL2291 PCR
´
´
GCGCGCCATATGTCTGAGCAAGATAACTACGGTT
saCOL2291
GCGCGCCTCGAGGTGAATGAAGTTATAACCAGCAG GCGCGCCATATGGCAGGACTTGCCACTATCGC
saCOL2581
GCGCGCCTCGAGATGAATGAAATTATATGAACCTGC
ScaC
ScaD BLAST
COL Puls4
saCOL2291 saCOL2291
bp ssaA2
COL
N
C5
Gene runner
ssaA2
2
1
M
1000bp 500bp
N C bp ssaA1 COL
saCOL2581 saCOL2581
ssaA1 C5
saCOL2291
PCR
SM0311
M
saCOL2581
saCOL2581 saCOL2291
saCOL2581
N C
saCOL2291
saCOL2581 saCOL2291
PCR
saCOL2291 saCOL2581 COL saCOL2291 saCOL2581 C saCOL2581 Kowsar Medical Journal
Vol. 16, No. 3, Fall 2011
saCOL2291
(PBP2a)
saCOL2581 saCOL2581 TATAAC (181-187) [6] GCTACA (178-184) [6]
CAT(40-42) [3]
A(331) T [5] G(112) A [6]
202bp [1]
30bp [1] T(514) A [5]
T(165) C [9] T(267) C [6] C(342) T [6] C(390) T [30] T(432) C [30] C(492) T [10] C(498) T [6]
saCOL2291
saCOL2581
saCOL2581 saCOL2581 Tyr+Asn#(61-62) [6] Ala+Tyr^(60-61) [6]
Asn(55) Asn [9] Asn(89) Asn [6] Pro(130) Ser [30] Ala(114) Val [6] Gly(166) Gly [6] Arg(164) Trp [10] Ala(144) Val [30]
Asn(111) Tyr [5]
His* [3]
Asp(38) Asn [6] -----
---Ala(172) Val [5]
^
saCOL
ScaC
ScaD
#
*
IsdB Sca (Sca) C
IsdB IsdA SdrE SdrD IsdB ScaD ScaC
saCOL2581 saCOL2291 COL N
COL saCOL2291 saCOL2291
COL DNA Pfu saCOL2581
saCOL2581
saCOL2291 saCOL2581
Huber LA, et al. Identification of vaccine candidate antigens of Staphylococcus aureus by serological proteome analysis. Proteomics. 2002;2(5):580-90. 9- Foster TJ. Potential for vaccination against infections caused by Staphylococcus aureus vaccine. Vaccine. 1991;9(4):221-7. 10- Hall AE, Domanski PJ, Patel PR, Vernachio JH, Syribeys PJ, Gorovits EL, et al. Characterization of a protective monoclonal antibody recognizing Staphylococcus aureus MSCRAMM protein clumping factor A. Infect Immun. 2003;71(12):6864-70. 11- Hu DL, Omoe K, Sasaki S, Sashinami H, Sakuraba H, Yokomizo Y, et al. Vaccination with nontoxic mutant toxic shock syndrome toxin 1 protects against Staphylococcus aureus infection. J Infect Dis. 2003;188(5):743-52. 12- Dumitrescu O, Tristan A, Meugnier H, Bes M, Gouy M, Etienne J, et al. Polymorphism of the Staphylococcus aureus Panton-valentine leukocidin genes and its possible link with the fitness of community-associated methicilin-resistant S. aureus. Infect Dis. 2008;198(5):792-4. 13- Blaiotta G, Fusco V, VonEiff C, Villani F, Becker K. Biotyping of entritoxigenic Staphylococcus aureus by entrotoxin gene cluster (egc) polymorphism. Appl Environ Mcrobial. 2006;72(9):6117-23. 14- Kim YK, Kim JS, Kim HS, Song W, Cho HC, Lee KM. Molecular typing of Staphylococcus aureus isolated from blood on the basis of coagulase gene polymorphism and toxin genes. Korean J Lab Med. 2008;28(4):286-92. 15- Silva ER, Boechat JU, Silva N. Coagulase gene polymorphism of Staphylococcus aureus isolated from goat mastitis in Brazilaien dairy herds. Lett Appl Microbial. 2006;42(1):30-4. 16- Shinefield H, Black S, Fattom A, Horwith G, Rasgon S, Ordonez J, et al. Use of a Staphylococcus aureus conjugate vaccine in patients receiving hemodialysis. N Engl J Med. 2002;346(7):491-6. 17- Senna JP, Roth DM. Protective immune response against methicillin-resistant Staphylococcus aureus in a murine model using a DNA vaccine approach. Vaccine. 2003;21(1920):2661-6. 18- Nelly AK, Desmond JC, Susan S, James C, Leslie DC, Tessie M, et al. A novel Staphylococcus aureus vaccine: Iron surface determinant B induces rapid antibody responses in Rhesus macaques and specific increased survival in a murine S. aureus sepsis model. Infec Immun. 2006;74(4):2215-23. 19- Yukiko K. Stranger J, Taeok B, Olaf S. Vaccine assembly from surface proteins of Staphylococcus aureus. Emil C, editor. New York: Rockefeller University Publication; 2006. 20- Dryla A, Prustomersky S, Gelbmann D, Hanner M, Beetinger E, Kocsis B. Comparison of antibody repertoires against Staphylococcus aureus in healthy individuals and in acutely infected patients. Clin Diagn Lab Immun. 2005;12(3):387-39. 21- Gill SR, Fouts DE, Archer GL, Mongodin EF, DeBoy RT, Ravel J, et al. Insights on evolution of virulence and resistance from the complete genome analysis of an early methicillinresistant Staphylococcus aureus strain and a biofilm-producing methicillin-resistant Staphylococcus epidermidis strain. J Bacteriol. 2005;187(7):2426-38. 22- Menbari SH, Pourmand MR, Shirazi MH, Mardani N. Cloning and sequencing of Sacol a novel gene from Staphylococcus aureus. Iran Med Microbial J. 2008;2(1):9-14.
Kowsar Medical Journal
Vol. 16, No. 3, Fall 2011
saCOL2581 saCOL2291
C
C
1- Charlebois ED, Perdreau-Remington F, Kreiswirth B, Bangsberg DR, Ciccarone D, Diep BA, et al. Origins of community strains of methicillin-resistant Staphylococcus aureus. Clin Infect Dis. 2004;39(1):47-54. 2- Dufour P, Gillet Y, Bes M, Lina G, Vandenesch F, Floret D, et al. Community-acquired methicillin-resistant Staphylococcus aureus infections in France: Emergence of a single clone that produces Panton-valentine leukocidin. Clin Infect Dis. 2002;35(7):819-24. 3- Centers for Disease Control and Prevention (CDC). Staphylococcus aureus resistant to vancomycin United States 2002. MMWR Morb Mortal Wkly Rep. 2002;51(26):565-7. 4- Lowy FD. Antimicrobial resistance: The example of Staphylococcus aureus. J Clin Investig. 2003;111(9):1265-73. 5- Josephson E, Hartford O, Obrien L, Patti JM, Foster T. Protection against experimental Staphylococcus aureus arthritis by vaccination with clumping factor A: A novel virulence determinant. J Infect Dis. 2001;184(12):1572-80. 6- Burine JP, Matthews RC, Carter T, Beaulieu E, Donohoe M, Chapman C, et al. Identification of an immunodominant ABC transporter in methicillin resistant Staphylococcus aureus infection. Infect Immun. 2000;68(6):3200-9. 7- Etz H, Min DB, Henics T, Dryla A, Winkler B, Triska C, et al. Identification of in vivo expressed vaccine candidate antigens from Staphylococcus aureus. Appl Biol Sci. 2002;99(10):6573-8. 8- Vytvytska O, Nagy E, Bluggel M, Meyer HE, Kurzbauer R,
