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重组IFN-α2a与HSV-tk/GCV系统协同作用增强对于卵巢癌细胞系SKOV3杀伤作用的体外研究
引用本文:程月鹃,潘凌亚,张毅.重组IFN-α2a与HSV-tk/GCV系统协同作用增强对于卵巢癌细胞系SKOV3杀伤作用的体外研究[J].中国肿瘤生物治疗杂志,2001,8(1):13-16.
作者姓名:程月鹃  潘凌亚  张毅
作者单位:[1]南京医科大学生化与分子生物学系,南京210029 [2]复旦大
摘    要: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BL21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BL21中诱导表在并纯化成功,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。

关 键 词:血管生成因子  血管生成抑制素  原核表达  多克隆抗体  制备
收稿时间:2000/12/11 0:00:00
修稿时间:2000年10月30

Prokaryotic Expression of Angiostatin and the Preparation of Polyclony Antibody
CHENG Yuejuan,PAN Lingya and ZHANG Yi.Prokaryotic Expression of Angiostatin and the Preparation of Polyclony Antibody[J].Chinese Journal of Cancer Biotherapy,2001,8(1):13-16.
Authors:CHENG Yuejuan  PAN Lingya and ZHANG Yi
Institution:Peking Union Medical College Hospital, Chinese Academy of Medical Science & Peking Union Medical College, Beijing 100730;Peking Union Medical College Hospital, Chinese Academy of Medical Science & Peking Union Medical College, Beijing 100730;Peking Union Medical College Hospital, Chinese Academy of Medical Science & Peking Union Medical College, Beijing 100730
Abstract: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.
Keywords:angiogenesis factor  angiostatin  prokaryotic expression  antibody
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