摘要
目的对螺旋藻脱辅基异藻蓝蛋白(apo-APC)的原核表达、蛋白纯化和抗肿瘤活性进行研究。方法通过基因工程技术,将异藻蓝蛋白基因(apc)克隆至pET28(a),转化JM109(DE3),构建其原核表达载体,并实现apo-APC蛋白在工程菌中高效表达;采用金属离子螯合亲和层析纯化目的蛋白,凝胶电泳分析其纯度,Western blot鉴定;以S180荷瘤小鼠为动物模型,分析apoAPC的抗肿瘤活性。结果通过基因工程技术获得apo-APC蛋白;动物实验显示apo-APC可抑制荷瘤小鼠体内S180肉瘤的生长,升高荷瘤小鼠的白细胞数量。结论螺旋藻藻胆蛋白的抗肿瘤活性至少有部分来自于其脱辅基蛋白。
Objective To investigate the prokaryotic expression, protein purification and anti-tumor activity of apo-allophycocyanin(apo-APC) in Spirulina. Methods Technique for gene engineering was used in the study. APC gene was subcloned into pET28(a), JM109(DE3) was translated and obtain high expression of apo-APC in engineering bacteria. The interest protein was purified by metal ion chelate affinity chromatography, its purity was analyzed by gel electrophoresis, the protein was identified by Western Blot. Sarcoma180 bearing mice were chosed as animal models to analyze the anti-tumor activity of apo-APC. Results The apo-APC could be obtained by technique for gene engineering. The animal experiment showed that apo-APC could inhibit the growth of Sarcoma180 in tumor-bearing mice, and increase leukocyte number of tumor-bearing mice. Conclusion The anti-tumor activity of phycobiliprotein in Spirulina may be partly due to its apoprotein at least.
出处
《福建中医药大学学报》
2014年第4期20-23,共4页
Journal of Fujian College of Traditional Chinese Medicine
基金
国家自然科学基金资助(81102844)
福建省教育厅资助(JA11134)
