Ju Chuan, Peng Jian-xin, Xu Bin, Wang Wei, Feng Zheng, Hu Wei
National Institute of Parasite Diseases, Chinese Centre for Disease Control and Prevention, Collaborating Center for Malaria, Schistosomiastis and Filariasis, Shanghai 200025, China.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2006 Oct;24(5):360-5.
To clone and express Schistosoma japonicum (Sj) calcyphosine gene, and purify the expressed protein.
The encoding sequence selected from Sj cDNA library was amplified by PCR. After subcloned into prokaryotic expression vector pET-28a, the expressed protein was purified with His -Tag affinity chromatography. Western blotting was used to detect the immunogenicity. The structure and functions of the protein were analyzed by bioinformatics method, and the phylogenetic tree of the protein was drawn.
The recombinant protein was specifically recognized by the Sj infected rabbit serum. The bioinformatics analysis showed 4 EF-hand domains. Besides, it was predicted that Sj calcyphosine contains two phosphorylation sites for protein kinase C, eight phosphorylation sites for casein kinase II and one N-myristoylation site. The Sj calcyphosine belonged to type-II calcyphosine.
The calcyphosine gene is a calcium-binding protein and might be a potential candidate for diagnosis, vaccine or drug target.
克隆并表达日本血吸虫（Sj）钙磷蛋白基因，并纯化表达的蛋白。
从Sj cDNA文库中选取编码序列，通过PCR进行扩增。亚克隆至原核表达载体pET-28a后，采用His标签亲和层析法纯化表达的蛋白。利用蛋白质免疫印迹法检测免疫原性。通过生物信息学方法分析该蛋白的结构和功能，并绘制该蛋白的系统发育树。
重组蛋白能被Sj感染兔血清特异性识别。生物信息学分析显示有4个EF手结构域。此外，预测Sj钙磷蛋白含有2个蛋白激酶C磷酸化位点、8个酪蛋白激酶II磷酸化位点和1个N-肉豆蔻酰化位点。Sj钙磷蛋白属于II型钙磷蛋白。
钙磷蛋白基因是一种钙结合蛋白，可能是诊断、疫苗或药物靶点的潜在候选物。
Zotero 插件