摘要
目的构建人绒毛膜促性腺激素(HCGβ)原核表达载体PG5-HCGβ,使HCGβ在大肠杆菌中获得高效表达。建立一条简单、快速、高效的纯化复性路线,获得具有生物活性的高纯度rHCGβ。方法以本室克隆的PCRII/HCGβ为模板,构建PG5-HCGβ,转化大肠杆菌BL21表达HCGβ。表达蛋白顺次用凝胶过滤和离子交换层析两步分离纯化。结果酶切及序列结果显示,PG5-HCGβ构建正确,HCGβ表达约占菌体总蛋白的20%。经过凝胶过滤和离子交换层析两步分离纯化,获得rHCGβ纯度达95%以上。复性的rHCGβ具有促进小鼠子宫生长的生物学活性。结论构建的PG5-HCGβ表达载体在大肠杆菌中获得高效表达,纯化复性的rHCGβ蛋白具有较高的生物学活性,为今后的rHCGβ生产打下了良好的基础。
Objective To construct PGS-HCG β Prokaryotic expression vectors of HCG β and express high-effectively recombinant HCG β. Based on HCG β in inclusion body in E.coli, a simple, fast and high-effective purification protocol was established. Methods PG5-rHCG β was constructed and transformed E.coli to express rHCG β using the previous cloned PCRII/HCG β as the template. The expression protein was orderly separated and purified using gel filtration and ion exchange chromatogram. Results PG5- rHCGβ was constructed successfully. The expression level was approximate 20% of the total protein. Purification of rHCG β was up to 95% after gel filtration and ion exchange chromatogram. The renatured rHCG β facilitate uterus growth of mouse. Conclusion PG5-HCG β can express high-effectively in E.coli, renatured rHCG β possess high biological activity.
出处
《江西医学检验》
2005年第6期523-525,共3页
Jiangxi Journal of Medical Laboratory Sciences
