山羊流产嗜衣原体OmpA基因重组质粒构建及对小鼠免疫效果研究

doi:10.16431/j.cnki.1671-7236.2016.11.012

Abstract

Abstract:

To explore the feasibility of entering clinical trials of the goat Chlamydophila abortus eukaryotic plasmids,Chlamydophila abortus OmpA gene was amplified by PCR and cloned into the eukaryotic expressing plasmid pcDNA3.1(+）to construction the recombinant vetor.After identification by PCR and restriction enzyme digestion,this vector was transfected into PK-15 cells and its expression were observed by fluorescent antibody test,the distribution of serum antibodies and plasmid were detected in mice after the immunization of the recombinant vector and molecular adjuvant which was single-stranded DNA of E.coli bacterial genome.The results showed that the recombinant plasmid pcDNA3.1-OmpA was successfully constructed after detecting by PCR,enzyme digestion and sequencing.The OmpA gene was effectively expressed in PK-15 cells.The anti-OmpA antibody levels of the pcDNA3.1-OmpA with molecular adjuvant group was significantly higher than that in pcDNA3.1-OmpA group and the inactivated vaccine group at 14 d after immunization（P<0.05).This levels showed an upward trend following numbers of immunization and times.The highest levels was at 35 d after immunization.Then the antibody titers were gradually decreased which still maintain a higher antibody levels than before.The pcDNA3.1-OmpA could be detected in the heart,liver,spleen,kidney,lung,brain,jejunum and leg muscle of mice on 21 d after immunization,and couldn't be detected in any organs at 49 d after immunization.The results above indicated that the the recombinant vetor based on OmpA gene of Chlamydophila abortus was successfully constructed in this experiment,after joining the molecular adjuvant could induce to a higher level of serum antibodies in mice.

Key words: Chlamydophila abortus; OmpA gene; molecular adjuvant; immune effect

CLC Number:

S855.9⁺9

LONG Chong-chong, DENG Wei-xi, MAO Yi-zhi, ZENG Gui-ying, WU Bo-tao, XIONG Chao-li, XU Mao-wen, XIA Peng, WEN Ming. Construction of Recombined Plasmid with OmpA Gene of Goat Chlamydophila abortus and Immunogenicity Effect in Mice[J]. , 2016, 43(11): 2886-2891.

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Construction of Recombined Plasmid with OmpA Gene of Goat Chlamydophila abortus and Immunogenicity Effect in Mice

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