The omp2 Gene of HPS-type Bacteria Cloning and Sequence Analysis Isolates from Sichuan Province

Lirui Li, Zexiao Yang, Yin Wang, Qiu Jin, Xulong Wu, Rongchang Zhang

Sichuan Agricultural University, Sichuan Laboratory of Animal Disease and Human Health, SLADHH, Ya'an ,China.
Tibet Changdu animal health and phytosanitary supervision, TCAHPS, Changdu, China.

DOI: 10.4236/eng.2012.410B040                

Abstract

In order to compare the homology and antigen of Haemophilus parasuis (HPS)4 outer membrane protein P2(omp2), we design a test with specific primers, using PCR amplification of isolates of Haemophilus parasuis from Sichuan Province(HP Sch2010), ompP2 gene will be cloned into the pGM-T vector, and transformed into E. coli DH5α. Identified by PCR and sequencing and analysis, the sequencing results showed that the published 4 HPS SW124 strains omp2 gene (1077bp), compared with the amplified 1086bp purpose fragment(containing omp2 genes), is relatively stable, with the nucleotide homology level 97% and amino acid homology level of 92.5%. The variable regions are mainly concentrated in the three base sequences: 40-65,110-156,180-202.

Keywords

Haemophilus parasuis, virulence genes, omp2

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Simone O,Carlos P.Haemophilus parasuis:new trends on diagnosis,epidemiology and control.Vet Microbiol. vol.99, pp.75-78, 2004
[2]	Oliveira A,Pina S,PerezSimo M, Marta Pérez-Simó,Simone Oliveira, Albert Bensaid.Virulence associated trimeric autotransporters of Haemophilus parasuis are antigenic proteins expressed in vivo.Vet Res. vol..41,pp.26, 2010.
[3]	M.Zhou,Y.Guo,J.Zhao.Identification and characterization of novel immunogenic outer membrane proteins of Haemophilus parasuis serovar 5.Vaccine.vol.27,pp.5271-5277,2009.
[4]	Haase EM,Campagnari A A,Sarwar J,Shero M,Wirth M,Cumming CU,et al.Strain- specific and immunodominant surface epitopes of the P2 porin protein of nontypeable Haemophilus influenzae.Infect Immun.. vol.59,pp.1278-1284,1991.
[5]	Hirokawa,Boon-Chieng,Mitaku.SOSUI classifica-tion,membrane protein two structure predic-tion.Bioinformatics,pp.378-379,1998
[6]	Romble IT,Sykes KF,Rayner S.ORF-FINDER:a vector for high-throughput gene identification.Gene. vol.282, pp.33-41, 2002.
[7]	Lillie B N,Hammerwueller J D,Macinnes J I, Mario Jacques, Anthony Hayes M.Porcine mannan-binding lectin A binds to Actinobacillus suis and Haemophilus parasuis.D Comp Immunol. vol.30,pp.954-965,2006．
[8]	V.A de la Puente Redondo,J Navas Méndez,N Garc??a del Blanco,N Ladrón Boronat,C.B Gutiérrez Mart??na, E.F Rodr??guez Ferri.Typing of Haemophilus parasuis strains by PCR-RFLP analysis of the tbpA gene.Vet Microbiol. vol..92, pp.253-262, 2003．
[9]	Lichtensteriger C A,Vimer E R.Purification and renaturation of membrane neuraminidase from Haemophilus parasuis.Vet Microbiol. vol..93,pp.79-87,2003．
[10]	Michael A M,Karen B R,Darrell O B,Crystal L L,Tracy L N, Susan L B,et al.Characterization and comparative analysis of the genes encoding Haemophilus parasuis outer membrane proteins P2 and P5.Journal of bacteriology. vol.191,pp.5988-6002,2009．
[11]	Martin de la,Fuente A J,Rodriguez-Freei E F,Martínez S,Tejerina F,Gutiérrez-Martín C B.Systemic antibody response in colostrums-deprived pigs experimentally infected with Haemophilus parasuis.Res Vet Sci. vol.86,pp.248-253,2009．
[12]	Tadjine M,Mittal K R,Bourdon S,Gottschalk M.Production and characterization of murine monoclonal antibodies against Haemophilus parasuis and study of their protection role in mice.Microbiol.. vol.150,pp.3935-3945,2004.
[13]	Mcvicker J,Tabatabai L,MurielV S.Haemophilus para-suis-novel proteins-hopes for vaccine.Respiratory diseases of livestock. vol.22,pp. 2005.
[14]	Nikolaj B,Thomas S P,Ramneek G, Steen G,S?ren B.Prediction of post-translational glycosylation and phosphorylation of proteins from the amino acid sequence. Proteomics. vol.4,pp.1633-1649,2004.
[15]	Mannm M,Jensen O N.Proteomic analysis of post-translational modifications.Nature Biotechnology. vol..21,pp.255-261,2003.