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PRL-3及其同源异形体重组杆粒的构建和鉴定 被引量：1

Construction of PRL-3 gene recombinant baculovirus vector
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摘要 目的构建PRL-3及其同源异形体重组杆状病毒表达载体,为下一步表达PRL-3及其同源异形体蛋白并从中草药活性成分中筛选出PRL-3抑制剂奠定基础。方法应用RT-PCR方法从高转移卵巢癌细胞HO-8910PM扩增PRL-3及其同源异形体编码序列,克隆到供体质粒pFastBac HTB,在DH10Bac菌中进行同源重组,经3种抗生素和蓝白斑筛选,得到Bacmid-HTB-PRL-3和Bacmid-HTB-PRL-3-variant,PCR鉴定其正确性。结果与结论成功构建了PRL-3及其同源异形体重组杆粒。 Objective To construct recombinant Bacmid of PRL-3 and its variant. Methods PRL-3 and its variant cDNA were amplified from total RNA of HO-8910PM by RT-PCR, and inserted into plasmid pFastBacHTB. The recombinant Bacmid of PRL-3 and its variant was screened by three antibiotics and Bluo-gal, confirmed by PCR. Results and Conclusion The re- combinant Bacmid of PRL-3 and its variant had been successfully constructed. It provides a basis for further study of expression of PRL-3 and its variant proteins and screening small molecule inhibitors from active components of Chinese traditional and herbal drugs.
作者 何太平 熊亮 莫丽儿 梁念慈
机构地区 广东医学院生物化学与分子生物学研究所 广东天然药物研究与开发重点实验室
出处 《广东医学院学报》 2008年第5期489-491,495,共4页 Journal of Guangdong Medical College
基金 广东省科技计划项目(No.2007B030702003) 广东省中医药局课题(No.1060044)
关键词 PRL-3 杆状病毒 基因重组 PRL-3 baculovirus gene recombination
分类号 Q784 [生物学—分子生物学]
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参考文献11

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二级参考文献15

  • 1KIM K A,SONG J S,JEE J,et al.Structure of human PRL-3,the phosphatase associated with cancer metastasis[J].FEBS Lett,2004,565(3):181-187.
  • 2KOZLOV G,CHENG J,LIEVRE C,et al.1H,13C and 15N resonance assignments of the human phosphatase PRL-3[J].J Biomol NMR,2002,24(2):169-170.
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  • 6ZENG Q,SI X,HORSTMANN H,et al.Prenylation-dependent association of protein tyrosine phosphatases PRL-1,-2,and -3 with the plasma membrane and the early endosome[J].J Biol Chem,2000,275(28):21444-21452.
  • 7LYON M A,DUCRUET A P,WIPF P,et al.Dual-specificity phosphatases as targets for antineoplastic agents[J].Nat Rev Drug Discov,2002,1:961-976.
  • 8SAHA S,BARDELLI A,BUCKHAULTS P,et al.A phosphatase associated with metastasis of colorectal cancer[J].Science,2001,294(5545):1343-1346.
  • 9MISKAD U A,SEMBA S,KATO H,et al.Expression of PRL-3 phosphatase in human gastric carcinomas:close correlation with invasion and metastasis[J].Pathobiology,2004,71(4):176-184.
  • 10WU X,ZENG H,ZHANG X,et al.Phosphatase of regenerating liver-3 promotes motility and metastasis of mouse melanoma cells[J].Am J Pathol,2004,164(6):2039-2054.

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广东医学院学报
2008年 第5期

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