血管生成抑素的原核表达及多克隆抗体的制备

Prokaryotic Expression of Angiostatin and the Preparation of Polyclony Antibody

目的 :在原核系统中表达并纯化人血管生成抑素 (angiostatin) ,制备鼠抗人血管生成抑素多克隆抗体。方法 :设计引物扩增angiostatin的cDNA ,然后亚克隆入原核表达载体pQE ,并转化入大肠杆菌BL2 1中诱导表达并纯化 ,再用纯化的人血管生成抑素免疫小鼠并制备多抗。结果 :通过重组质粒酶切和测序分析等方法 ,筛选出重组阳性克隆 ,转化入大肠杆菌BL2 1中诱导表达并纯化成功 ,再利用纯化的人血管生成抑素制备成功鼠抗人血管生成抑素多克隆抗体。结论 :表达产物及多克隆抗体为下阶段深入研究提供了重要的实验材料。

Objective:Express and purify the recombant protein in prokaryotic system, preparing the mouse anti human angiostatin polyclony antibody.Methods: The cDNA of angiostatin was amplified by PCR and was subcloned into vector pQE-30, and transformed into the E.coli BL21(DE3). At last angiostatin was purified and used to immune the mouse for preparing polyclony antibody .Results: The recombinant clones were picked out by the methods of restrictional enzyme cut and sequence analysis. Recombinant angiostatin was highly expressed when the strain was induced with 1mmol/L IPTG. Angiostatin was successfully purified and the polyclony antibody was also successfully prepared. Conclusion: The recombinant protein and the polyclony antibody can be used in further studies.