禽多杀性巴氏杆菌OmpADNA疫苗免疫效果地研究.pdfVIP

454 201127 doi: 10. 3969/ j. issn. 10004 4X. 2011.05.017 OmpA DNA ( 河南科技大学, 洛阳471003) S855. 1 A 1000484X( 2011) [ ] : OmpA DNA : PCR ompa , pcDNA3. 1( + ) , pOMPA, SP2/ 0 , RTPCR : pOMPA pCDNA3. 1( + ) PBS , 15 4 , , , 1 / ELISA ,MTT , ELISA IFN~ , , : pOMPA , DNA , ( P 0.01) , DNA (P 0.05) ( Omps) , SI pCDNA3. 1( + ) PBS (P 0.01) , IFN( P 0. 01) DNA 60% 73. 3% : OmpA DNA , , [ ] ; OmpA DNA ; ; The immune efficacy of OmpADNA vaccine agains avian pas eurella mul ocida GONG Qiang , WANG ShuaiTao, QIN CuiL i, NI UM ingFu, CHENG Ming, HOU YuZe, ZHANG YongFa, S UN X iao Fei. H enan Unive sity of Science Technology , Luoyang 471003, China [Abs rac ] Objec ive:To research on protective immunity of OmpADNA vaccine against fowl cholera in chickens.Me hods :The ompa gene fragment amplified by PCR from avian Pasteurella multocidawas cloned into the eukaryotic expression vector pcDNA3. 1( + ) , and the re combinant plasmid pOMPA was constructed.Then the recombinant plasmidwas transfected into SP2/ 0 cells in vitro.The transcription and ex pression of target gene were analyzed by RTPCR and indirect immunofluorescence. Four groups of chickens named pOMPA, attenuated live vac cine, pCDNA3. 1( + ) and PBS were vaccinated with the DNA vaccine, attenuated live vaccine, control vector and PBS respectively.The serum antibodies were detected by indirect ELISA.The peripheral blood lymphocyte ( PBLC) proliferation and secreted of PBLC were tested by MTT. The immunized chickens were challengedwith virulent of avian Pasteurella multocida on week 2 post the third immunization, and the protection rate was counted. Resul s:RTPCR and indirect immunofluorescence showed that the ompa gene could be transfected into SP2/ 0 cells in vitro and the target proteinwas expressed. Indirect ELISA showed that the levels of anti