摘要
目的 构建在E .coli中表达人La多肽的质粒。方法以人脾cDNA为模板 ,通过PCR获得La多肽 (全长 )的DNA片段。将此片段利用双酶切定向克隆到MBP(麦芽糖结合蛋白 )融合系统的载体pMALTM c中 ,表达可溶性融合蛋白 ,并通过亲和层析予以纯化。结果免疫印迹实验 (IBT)证明 ,表达的融合蛋白具有La抗原多肽的特异性 ,而敏感性超过生化制备的ENA制品。
Aim To construct a vector expressing human La polypeptide in E.coli. Methods The fulllength DNA fragment encoding La polypeptide was obtained by PCR from human spleen cDNA. The La DNA fragment was cloned into plasmid pMALc(a maltosebinding protein, MBP fusion vector) and expressed in E.coli. The soluble MBPLa fusion proteins were purified by affinity chromatography using amylose resin. Results Western blot showed that the fusion protein could react with antiLa antibody as the natural La polypeptide, and the sensitivity was higher than natural antigen. Conclusion The fusion La polypeptide has the immunoidentity with natural La polypeptide.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
2000年第3期207-210,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
河北省自然科学基金!No.396062
