A quantitative assay method of Toxoplasma gondii HSP70 mRNA by quantitative competitive-reverse transcriptase-PCR

descriptionPublicationkeyboard_double_arrow_right Article 01 Mar 2004 English Publisher:Elsevier BVJournal:Parasitology International, volume 53, pages 49-58 (issn: 1383-5769,

Authors: Lian-Xun, Piao; Fumie, Aosai; Mei, Chen; Hao, Fang; Hye-Seong, Mun; Kazumi, Norose; Akihiko, Yano;

doi: 10.1016/j.parint.2003.11.001

pmid: 14984835

A quantitative assay method of Toxoplasma gondii HSP70 mRNA by quantitative competitive-reverse transcriptase-PCR

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Abstract

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.

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Chiba University
Japan

Keywords

Mice, Knockout, Mice, Inbred BALB C, Virulence, Reverse Transcriptase Polymerase Chain Reaction, Protozoan Proteins, Interferon-gamma, Mice, Toxoplasmosis, Animal, Liver Function Tests, Animals, HSP70 Heat-Shock Proteins, RNA, Messenger, Toxoplasma

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