A quantitative assay method of Toxoplasma gondii HSP70 mRNA by quantitative competitive-reverse transcriptase-PCR.
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.
Department of Infection and Host Defense, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-Ku, Chiba 260-8670, Japan.
Lian-Xun
LX
Piao
Fumie
F
Aosai
Mei
M
Chen
Hao
H
Fang
Hye-Seong
HS
Mun
Kazumi
K
Norose
Akihiko
A
Yano
D000818:Animals; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D007371:Interferon-gamma / Q000235:genetics; D008111:Liver Function Tests; D051379:Mice; D008807:Mice, Inbred BALB C; D018345:Mice, Knockout; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014122:Toxoplasma / Q000235:genetics / Q000254:growth & development / Q000378:metabolism / Q000472:pathogenicity*; D014124:Toxoplasmosis, Animal / Q000401:mortality / Q000469:parasitology; D014774:Virulence
D018840:HSP70 Heat-Shock Proteins; D015800:Protozoan Proteins; D012333:RNA, Messenger; D007371:Interferon-gamma