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  厄爾多思2 2007-04-28 00:00:00

  The M gene is one of infectious bronchitis virus important immunity original genes， the M gene possibly has the important antigen decision bunch， also has highly the conservative nature in the evolution process. This research selects the M gene of IBV ， on the foundation of researching the IBV structure protein gene， expands the PCR increases introduces the BamH I enzyme in the beginnings and ends to cut the position spot， cuts the insertion after the enzyme "the suicide" DNA vaccine carrier pSCA1 CMV the IE start under clone position spot， constructed "the suicide" the DNA vaccine to express material particle pSCA1-M. By the way of colony PCR， the enzyme cut and measured the foreword appraisal goal gene succeeds the clone to carrier pSCA1， also the direction is correct. Separately takes the isometric reorganization material particle pSCA1-M， conventional IBV M gene DNA vaccine pcDNAM and the spatial carrier pSCA1 extension dyes the Vero cell. After 48h， by the indirect immunity fluorescence experiment observation we discovery that pSCA1-M as well as the pcDNAM extension dyes the group 細(xì)胞漿 obviously specificity bright green fluorescence， the pSCA1 spatial carrier group does not have the fluorescence is the negative. This confirmed "the suicide" DNA vaccine carries can express the M gene successfully. But the pSCA1-M group institute luminescence cell number are less than the pcDNAM group which shows "the suicide" DNA vaccine extension dyeing rate to be lower than the conventional DNA vaccine. In order to appraise IBV "the suicide" DNA vaccine immunity effect， we use constructly "the suicide" DNA vaccine pSCA1-M， conventional DNA vaccine pcDNAM， spatial carrier pSCA1 and the IBV 滅活 vaccine separately immunity non- immunity health young chicken， and supposes the comparison group to inject the isometric PBS fluid. Since every week after immunization group chickens blood serum separation sustained five weeks. Using FACS and indirect ELISA test regarding 宿主外周 T lymphocyte subpopulations CD 3 +， CD4 +， CD8 + and specificity of serum IgG anti-IBV dynamic change detection separately， after the fifth week take the IBV strains strong attacking drug protection experiment. The indirect ELISA experiment examination result indicated that， we construct "the suicide" DNA vaccine be able to stimulate the organism to produce in view of the IBV specificity blood serum immune body. The third week after the immunity rise the peak value， afterwards starts to drop， its immune body level quite to conventional IBV and the DNA vaccine， is lower than the 滅活 vaccine. The T lymphocyte subgroup dynamic change result showed that， "the suicide" DNA vaccine pSCA1-M group and CD3+， CD4+， CD8+ each T lymphocyte quantity is higher than others. It displays difference remarkable (P＜0.01 or P＜0.05). But it has an obvious drop about at the 4th week， and slightly afterwards. It indicated that "the suicide" DNA vaccine can induce the organism to produce higher cellular immunity than the conventional DNA vaccine and 滅活 vaccine ， but rapid drop along with the death of 宿主 cell. The attacks poisonous protection test result statistics that， the pSCA1-M group chicken disease incidence's rate is 12%， the protection's rate is 50%; The pcDNAM group chicken disease incidence's rate is 13%， the protection's rate is 40%， which is slightly lower than pSCA1-M; The spatial carrier group disease incidence's rate (75%) ＜ the comparison group (85%)， two groups of the attacks poisonous protection's rate are in the same as 25%; 滅活苗組無雞 only arises the death. 對強(qiáng)毒的抵抗力單基因的“自殺性”DNA疫苗與常規(guī)DNA疫苗相當(dāng)（40%）＜滅活苗（）。 This research uses the IBV M gene to construct "the suicide" DNA vaccine to immunity the chicken， and carries on the appraisal to its immunity effect. It has not been reported at home and in the world . It can provide the foundation material and the scientific basis for researching IBV "the suicide"DNA vaccine. 有句不會翻譯~

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