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人血管生成素cDNA的克隆与表达 被引量：4

Cloning and Expression of Human Angiogenin cDNA in E.coli
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摘要 血管生成素 ( angiogenin,ANG)广泛存在于多种肿瘤组织中 ,在肿瘤发生的不同阶段刺激新生血管的形成 .利用 RT- PCR方法从培养的人肺癌细胞系 A549扩增得到了 ANG c DNA片段 ,测序正确后克隆入融合表达载体 p RSETB中 ,构建了原核表达菌株 .经 IPTG诱导 ,表达了 N端融合His6的 ANG融合蛋白 ,表达量占菌体总蛋白的 1 0 % ,纯化后的血管生成素体外能够有效地刺激鸡胚绒毛尿囊膜的血管形成 . The cDNA fragment encoding hANG was amplified by reverse transcription polymerase chain reaction method from the human lung cancer cell line A549 and inserted into the prokaryotic fusion expression vector pRSETB.The SDS PAGE showed that E.coli BL21 DE3(pLysS)containing pRANG expressed a His6 fusion protein.The expression level of the rhANG was about 10% of the total cell proteins.Biological assay result indicated that rhANG could induce the new blood vessel formation on CAM in vitro.
作者 杨辉 张英起 颜真 韩苇 药立波 苏成芝
机构地区 第四军医大学生物技术中心
出处 《中国生物化学与分子生物学报》 CAS CSCD 2000年第4期473-477,共5页 Chinese Journal of Biochemistry and Molecular Biology
关键词 血管成生 RT-PCR 基因克隆 基因表达 Angiogenin,RT PCR,Gene expression
分类号 Q78 [生物学—分子生物学] R730 [医药卫生—肿瘤]
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参考文献13

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同被引文献30

  • 1隋广超,胡美浩.影响大肠杆菌中外源基因表达的因素[J].生物化学与生物物理进展,1994,21(2):128-132. 被引量：55
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  • 5Denefle P, Kovarik S, Guitton JD, et al. Chemical synthesis of a gene coding for human angiogenin, its expression in Escherichia coli and conversion of the product into its active form[J]. Gene,1987,56(1):61-70.
  • 6Leland PA, Staniszewski KE, Park C, et al. The ribonucleolytic activity of angiogenin [J]. Biochem,2002,41(4) :1343-1350.
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中国生物化学与分子生物学报
2000年 第4期

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