论文部分内容阅读
为观察恶性疟原虫 FCC- 1/ HN株 pc DNA3- EBA175 / HRP 重组质粒经肌肉途径接种后在宿主体内的表达。BAL B/ c小鼠分为 3组 ,1组 (实验组 )经后腿股四头肌注射重组质粒 pc DNA3- EBA175 / HRP 10 0 μg;2组 (实验对照组 )注射空白质粒 pc DNA3 10 0 μg;3组 (正常对照组 )注射 PBS(p H7.4) 10 0 μl。每隔 2 0 d加强注射 ,于第 1次接种后 2 0 d和第 2次接种后 10 d取小鼠双侧股四头肌免疫组化染色 ,于第 3次接种后 40 d取小鼠各组织作聚合酶链反应 (PCR)检测目的基因。结果显示 ,(1) 1组经重组质粒免疫小鼠血清、恶性疟原虫抗原免疫小鼠血清、恶性疟原虫人工合成九肽 HRP 单克隆抗体及抗恶性疟原虫全虫抗原单克隆抗体作用 ,免疫组化染色后 ,在注射部位肌肉组织的肌膜、肌间隙处见到褐色分泌颗粒 ;1组未注射侧肌肉组织、2组及 3组均未见褐色颗粒。 (2 )采用PCR从 1组肌肉、心脏、肝脏、肺脏和肾脏组织扩增出目的基因 EBA175和 HRP ,2组及 3组相应组织均未扩增出目的基因片段。本研究为疟疾多基因 DNA疫苗的研制提供了体内表达的依据
To observe the expression of Plasmodium falciparum FCC-1 / HN strain pcDNA3-EBA175 / HRP recombinant plasmid in the host after intramuscular inoculation. BALB / c mice were divided into three groups. Group 1 (experimental group) was injected with pcDNA3-EBA175 / HRP pcDNA3-EBA175 / HRP into the quadriceps femoris muscle of the hind legs. Two groups (experimental control group) 0 μg; 3 groups (normal control group) injected PBS (p H7.4) 10 0 μl. The injection was boosted every 20 days. The bilateral quadriceps femoris immunohistochemical staining was performed on the day 20 after the first vaccination and 10 days after the second vaccination, and the mice were taken 40 days after the third vaccination Each organization detected the target gene by polymerase chain reaction (PCR). The results showed that: (1) 1 group of mice immunized with recombinant plasmid-immunized mice serum, Plasmodium falciparum antigen-immunized mice, artificial synthetic nonapeptide HRP monoclonal antibody of Plasmodium falciparum and alloplasma antigen of Plasmodium falciparum, immunized After histochemical staining, brown secreted granules were found at the muscle membrane and intramuscular space at the injection site. In group 1, uninjected muscle tissue was observed. No brown granules were observed in groups 2 and 3. (2) The target gene EBA175 and HRP were amplified from the muscle, heart, liver, lung and kidney tissue of 1 group by PCR, and the corresponding gene fragments were not amplified in 2, 3 and 3 groups. This study provided the basis for the in vivo expression of malaria multi-gene DNA vaccine