弓形虫ROP5基因真核表达载体的构建及其免疫保护性的研究

摘 要：【目的】构建弓形虫pCDNA-ROP5真核表达载体，观察弓形虫ROP5基因在小鼠体内诱导的免疫反应。【方法】克隆弓形虫ROP5基因，将其插入真核表达载体pCDNA-3.1中构建重组质粒pCDNA-ROP5，脂质体法转染Hela细胞，肌注免疫昆明小鼠，测定小鼠血清中的IgG，观察小鼠受攻击感染后的存活时间。【结果】真核表达载体构建成功，实验组pCDNA-ROP5免疫的小鼠与对照组相比，实验组小鼠产生了明显的体液免疫应答；攻击感染试验中，实验组的小鼠比对照组的存活时间有明显延长。【结论】构建的真核表达重组质粒pCDNA-ROP5对抗击弓形虫攻击感染能够激发一定的免疫保护性。

Construction of eukaryotic expression vector of Toxoplasma gondii ROP5 gene and its immunoprotective effect

Abstract【Objective】To construct eukaryotic expression pCDNA-ROP5 and evaluated the immune responses it induced in Kunming mice.【Methods】ROP5 gene fragment was amplified by PCR and insert into pCDNA-3.1 vector to form the recombinant plasmid pCDNA-ROP5;Hela cells were transfected by plasmid pCDNA-ROP5 with lipofection strategy;plasmid pCDNA-ROP5, pCDNA-3.1 and NS were injected into Kunming mice muscle three times, antibody IgG level were measured in each group; invested the survival time after challenged by T. gondi【Results】The results showed that the ROP5 has been successfully expressed in Hela cells and developed a high level of specific antibody response, Survival time of ROP5 immune mice was prolonged when infected by T.gondii.【Conclusion】ROP5 could trigger immmuno-protection against T. gondii challenge.