zs株弓形虫P22基因片段在巨噬细胞中的表达

Expression of the P22 gene fragment of ZS strain of Toxoplasma gondii in macrophages

目的 克隆表达ZS株弓形虫表膜抗原P2 2编码基因的巨噬细胞株。方法 扩增P2 2编码基因ORF的全长 5 6 1bp片段 ,经PstI、XbaI双酶切后 ,定向克隆于质粒 pBudCE 4 .1,构建真核重组表达质粒 pBudCE 4 .1/P2 2 ,通过脂质体介导转染入小鼠巨噬细胞RAW2 6 4 .7,以RT PCR方法鉴定目的基因在巨噬细胞中的表达。结果 成功获取pBudCE 4 .1/P2 2转染的巨噬细胞阳性克隆 ,并证实P2 2 mRNA在阳性克隆细胞中的表达 ,为P2 2蛋白对巨噬细胞的免疫调节功能作用的研究奠定基础。

To clone the positive clones of macrophages expressing the P22 gene fragment of ZS strain of Toxoplasma gondii, the whole length of 561 bp fragment coding gene ORF of the P22 gene was amplified by PCR and digested by PstI and XbaI. The amplified fragment was cloned into pBudCE4.1 to construct the recombinant plasmid pBudCE4.1/P22, and then the murine macrophages were transfected with this gene fragment by using lipofectamine. The expression of the P22 mRNA in the transfected macrophages was identified by RT PCR. By using the methods mentioned above, the positive clones of the macrophages transfected with pBudCE4.1/P22 were successfully isolated.