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FHL家族的克隆及其融合蛋白的纯化 被引量：2

Cloning of FHL Family and Purification of FHL Fusion Proteins
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摘要 目的:纯化FHL家族(FHL1～FHL5)融合表达蛋白。方法:用PCR方法从乳腺文库和卵巢文库中扩增FHL1～FHL5编码序列,将其以正确相位与pGEX-KG载体中的GST编码序列融合,将重组质粒分别转化E.coliDH5α后,用谷胱甘肽-Sepharose4B纯化融合蛋白,并用Westernblot检测融合蛋白的表达。结果:构建得到FHL1～FHL5的融合表达载体,West-ernblot检测表明GST-FHL1～GST-FHL5共5种融合蛋白均成功表达,并纯化得到融合蛋白。结论:成功克隆FHL1～FHL5基因,并得到纯化的融合蛋白。 Objective： To purify the fusion protein of FHL family （FHL1-FHL5）. Methods： The coding sequences of FHL1-FHL5 were amplified by PCR and fused in frame with the coding region of GST in the pGEX-KG vector to generate the pGST-FHLI-pGST-FHL5 recombinant plasmids.The fusion proteins GST-FHL1-GST-FHL5 were expressed in E. coli DH5α and purified by Glutathione-Sepharose 4B affinity chromatography. The fusion proteins were characterized by Western blot. Results： The pGST-FHLI-pGST-FHL5 recombinant plasmids and purified fusion proteins were successfully obtained. Western blot analysis showed that the fusion proteins were expressed. Conclusion： FHL1-FHL5 were successfully cloned and their fusion proteins were purified.
作者 丁丽华 方言 严景华 袁斌 韩聚强 吕秋军 叶棋浓
机构地区 军事医学科学院生物工程研究所 军事医学科学院放射医学研究所
出处 《生物技术通讯》 CAS 2006年第2期174-176,共3页 Letters in Biotechnology
基金 国家自然科学基金项目(30530320 30500091 30428012)
关键词 FHL 克隆 融合蛋白 FHL cloning fusion protein
分类号 Q786 [生物学—分子生物学] Q503 [生物学—生物化学]
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参考文献21

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同被引文献13

  • 1丁丽华,袁斌,严景华,韩聚强,姜艳超,李杰之,叶棋浓.FHL1抗体的制备与特异性鉴定[J].细胞与分子免疫学杂志,2006,22(6):804-806. 被引量：4
  • 2Brown S,Biben C,Ooms LM,et al.The cardiac expression of striated muscle LIM protein 1 (SLIM1) is restricted to the outflow tract of the developing heart[J].J Mol Cell Cardiol,1999,31(4):837-843.
  • 3Chu PH,Ruiz-Lozano P,Zhou Q,et al.Expression patterns of FHL/SLIM family members suggest important functional roles in skeletal muscle and cardiovascular system[J].Mech Dev,2000,95(1-2):259-265.
  • 4Fimia G,De Cesare D,Sassone-Corsi P.CBP-independent activation of CREM and CREB by the LIM-only protein ACT[J].Nature,1999,398(6723):165-169.
  • 5Lee SM,Li HY,Ng On EK,et al.Characterization of a brain-specific nuclear LIM domain protein (FHL1B) which is an alternatively spliced variant of FHL1[J].Gene,1999,237(1):253-263.
  • 6Yang J,Moravec CS,Sussman MA,et al.Decreased SLIM1 expression and increased gelsolin expression in failing human hearts measured by high-density oligonucleotide arrays[J].Circulation,2000,102(25):3046-3052.
  • 7Ng On EK,Lee SM,Li HY,et al.Characterization of tissue-specific LIM domain protein (FHL1C) which is an alternatively spliced isoform of a human LIM-only protein (FHL1)[J].J Cell Biochem,2001,82(1):1-10.
  • 8Fimia GM,Cesare DD,Corsi PS.A Family of LIM-Only Transcriptional Coactivators:Tissue-Specific Expression and Selective Activation of CREB and CREM[J].Mol Cell Biol,2000,20(22):8613-8622.
  • 9Sanchez-Garcia I,Rabbitts TH.The LIM domain:a new structural motif found in zinc-finger-like proteins[J].Trends Genet,1994,10:315.
  • 10Brown S,Biben C,Ooms LM,et al.The cardiac expression of striated muscle LIM protein 1 (SLIM1) is restricted to the outflow tract of the developing heart[J].J Mol Cell Cardiol,1999,31(4):837-843.

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生物技术通讯
2006年 第2期

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