Cloning,fusion expression and identification of thioredoxin encoding gene from Toxoplasma gondii
10.16250/j.32.1374.2016077
- VernacularTitle:刚地弓形虫硫氧还蛋白基因的克隆表达及鉴定
- Author:
Zigang ZHANG
;
Xiaomei CHEN
;
Danhua SU
;
Yuan LIU
;
Tao FU
;
Jiamiao DUANMU
;
Liang WU
;
Xugan JIANG
;
Shengxia CHEN
;
Jianping CAO
- Publication Type:Journal Article
- Keywords:
Toxoplasma gondii;
Thioredoxin;
Cloning;
Prokaryotic expression;
Identification;
Polyclonal antibody
- From:
Chinese Journal of Schistosomiasis Control
2016;28(3):289-292
- CountryChina
- Language:Chinese
-
Abstract:
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.
