Piao Lian-Xun, Aosai Fumie, Chen Mei, Fang Hao, Mun Hye-Seong, Norose Kazumi, Yano Akihiko
Department of Infection and Host Defense, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-Ku, Chiba 260-8670, Japan.
Parasitol Int. 2004 Mar;53(1):49-58. doi: 10.1016/j.parint.2003.11.001.
Toxoplasma gondii (T. gondii)-derived heat shock protein 70 (T.g.HSP70) has been identified as a virulent molecule expressing only in T. gondii tachyzoites during lethal acute infection. Therefore, it is of importance to determine the expression of T.g.HSP70 mRNA in a quantitative manner for analysis of virulence of T. gondii in tissues. We have constructed a competitor T.g.HSP70 and have successfully established a quantitative competitive-reverse transcriptase-polymerase chain reaction (QC-RT-PCR) targeting T.g.HSP70 gene. By using the established QC-RT-PCR method, we have demonstrated that the copy number of T.g.HSP70 mRNA per T. gondii tachyzoite was highest in the lung among the organs examined in interferon-gamma knockout (GKO) mice.
刚地弓形虫（T. gondii）衍生的热休克蛋白70（T.g.HSP70）已被鉴定为一种仅在致死性急性感染期间于刚地弓形虫速殖子中表达的毒力分子。因此，以定量方式确定T.g.HSP70 mRNA的表达对于分析刚地弓形虫在组织中的毒力很重要。我们构建了一个竞争性T.g.HSP70，并成功建立了针对T.g.HSP70基因的定量竞争性逆转录-聚合酶链反应（QC-RT-PCR）。通过使用已建立的QC-RT-PCR方法，我们证明在干扰素-γ基因敲除（GKO）小鼠所检测的器官中，每个刚地弓形虫速殖子的T.g.HSP70 mRNA拷贝数在肺中最高。
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