摘要
目的体外制备华支睾吸虫CsTC基因工程蛋白,并进行免疫学研究。方法用组氨酸标签亲和纯化试剂盒纯化重组蛋白CsTC。将CsTC蛋白免疫BALB/c小鼠,制备其多抗血清。酶联免疫吸附(ELISA)检测抗血清抗体滴度,蛋白质印迹法(Western blotting)鉴定其抗血清的特异性。结果纯化蛋白CsTC的浓度为(0.78±0.03)mg/ml,纯度达90%。间接ELISA结果显示,免疫过程中抗体滴度持续上升。Western-blot结果显示,免疫小鼠血清具有抗CsTC特异性。结论本研究制备的CsTC基因工程蛋白和抗CsTC抗血清可较好地用于后续CsTC的基因功能鉴定。
The transcriptional coactivator gene product recombinant protein CsTC of Clonorchis sinensis was prepared in vitro by purification of this protein with His Bind Purification Kit and BALB/c mice were immunized with this protein to prepare its polyclonal antibody. ELISA assay was used to detect the antibody titer of antisera and Western blot assay was employed to detect the specificity of antisera. It was found that the concentration of the purified CsTC protein was （0.78±0.03） mg/ml and its purity was 90%. As demonstrated by ELISA, the titer of antibodies increased steadily in the course of immunization. Western blot assay displayed that CsTC protein had high anti-CsTC specificity. These results suggest that the CsTC protein prepared via genetic engineering can be used for the functional identification of the gene encoding CsTC.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2007年第3期259-261,268,共4页
Chinese Journal of Zoonoses
基金
广东省自然科学基金团队项目(No.20003026)
"211"重点学科基金(No.98169)
广东药学院2004年度科研项目(广药科〔2005〕02号)资助
