Zhongguo xue xi chong bing fang zhi za zhi = Chinese journal of schistosomiasis control · 2017
[Preliminary research on prokaryotic expression and immune protection of triosephosphate isomerase of Toxoplasma gondii].
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Shuang Shen, Xu-Ren Yin, Li-Jun Song, Jie Wang, Xue-Dan Ke, Wei Zhou, Chuan-Xin Yu
Аннотация
OBJECTIVE: To study the prokaryotic expression and immune protection of triosephosphate isomerase (TPI) of Toxoplasma gondii in mice.
METHODS: Total RNA was extracted from toxoplasma tachyzoites, and TPI fragment was amplified by PCR and cloned into the prokaryotic expression vector pET-28a (+). The target protein was induced with IPTG and purified by Ni-NTA affinity chromatography. The mice were immunized 4 times by emulsified TPI with adjuvant, and the last time was the strengthen immunization. At the same time, an adjuvant group and a normal group were set as controls. The blood samples were got from the tail vein of the mice, and the serum antibody titres were detected. All the mice were challenged with 400 toxoplasma tachyzoites to observe the survival time.
RESULTS: The TPI gene was amplified from T. gondii cDNA by PCR. The recombinant vector TPI/pET-28a (+) was usefully constructed, and the TPI protein was expressed and purified. The serum antibody titre could be more than 100 thousand. After infected with toxoplasma tachyzoites, the survival time of the mice in the experimental group was longer than that of the mice in the control groups.
CONCLUSIONS: The TPI protein of T. gondii could trigger the immunoprotection against T. gondii challenge in the mice.
METHODS: Total RNA was extracted from toxoplasma tachyzoites, and TPI fragment was amplified by PCR and cloned into the prokaryotic expression vector pET-28a (+). The target protein was induced with IPTG and purified by Ni-NTA affinity chromatography. The mice were immunized 4 times by emulsified TPI with adjuvant, and the last time was the strengthen immunization. At the same time, an adjuvant group and a normal group were set as controls. The blood samples were got from the tail vein of the mice, and the serum antibody titres were detected. All the mice were challenged with 400 toxoplasma tachyzoites to observe the survival time.
RESULTS: The TPI gene was amplified from T. gondii cDNA by PCR. The recombinant vector TPI/pET-28a (+) was usefully constructed, and the TPI protein was expressed and purified. The serum antibody titre could be more than 100 thousand. After infected with toxoplasma tachyzoites, the survival time of the mice in the experimental group was longer than that of the mice in the control groups.
CONCLUSIONS: The TPI protein of T. gondii could trigger the immunoprotection against T. gondii challenge in the mice.