环形泰勒虫spm-N蛋白的原核表达及鉴定

Prokaryotic expression and identification of spm-N protein of Theileria annulata

子孢子与裂殖体2(spm2)是环形泰勒虫重要的表面抗原。为了深入研究环形泰勒虫spm2主要抗原区域spm-N蛋白的功能,利用大肠杆菌表达系统表达重组spm-N蛋白并进行纯化及鉴定。通过序列设计合成特异性引物,利用PCR扩增环形泰勒虫spm-N基因片段,将目的片段定向克隆至表达载体pET30a(+),转化大肠杆菌BL21(DE3)表达菌,用IPTG诱导表达重组蛋白。结果显示:重组spm-N蛋白以可溶性的形式表达,获得了较高纯度的spm-N蛋白,且该蛋白的反应原性和特异性均较好。本研究成功表达了重组spm-N蛋白,为深入揭示spm-N蛋白的功能提供了依据。

The establishment of reliable and convenient diagnostic methods is the key to preventing and controlling epidemic diseases;and selection of reaction with originality and specificity,and of low cost and applicability in mass production of antigen,is the precondition to establish diagnosis methods.This study was to characterize the spm-N protein of Theileria annulata.The Prokaryotic expression of the spm-N gene of the Theileria annulata was investigated and purified proteins were obtained and identified.The spm-N gene was amplified by PCR and inserted into plasmid pET30a(+)to construct a recombinant plasmid pET30a-spm-N which was transformed into E.coli BL21 and whose expression was induced with IPTG.The results showed that pET30a-spm-N was expressed in a soluble form and was purified successfully.The recombinant protein also had good repeatability and sensitivity.In conclusion,the pET30a-spm-N of Theileria annulata was expressed on E.coli,which laid a foundation for research on the function of spm-N.