1.Cloning,fusion expression and identification of thioredoxin encoding gene from Toxoplasma gondii
Zigang ZHANG ; Xiaomei CHEN ; Danhua SU ; Yuan LIU ; Tao FU ; Jiamiao DUANMU ; Liang WU ; Xugan JIANG ; Shengxia CHEN ; Jianping CAO
Chinese Journal of Schistosomiasis Control 2016;28(3):289-292
Objective To clone and express the thioredoxin(Trx)from RH strain tachyzoites of Toxoplasma gondii,estab?lish the prokaryotic expression vector and purify the recombinant protein,then produce the polyclonal anti?Trx antibody in rab?bits. Methods Trx fragment was amplified by PCR and cloned into the pET?28a(+)vector,and the recombinant protein was in?duced with IPTG and purified by Ni?NTA affinity chromatography. The polyclonal antibody specificity was detected by Western blotting. Results The trx gene was amplified from T. gondii cDNA by PCR. The recombinant plasmid trx/pET?28a(+)was use?fully constructed,and the recombinant TRX protein was expressed and purified. The TRX polyclonal antibody was also ob?tained. The specific band of TRX was detected by Western blotting. Conclusion Western blotting can detect the specificity of polyclonal anti?Trx antibody,which will facilitate the biological functions of Trx.
Result Analysis
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