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荧光表达载体Podocin过表达对HEK293细胞中CD2AP分布的影响 被引量：1

Construction of expression vector for podocin and its effect of transfection on CD2AP distribution in HEK293 cells
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摘要 目的:构建podocin荧光表达载体,并观察其转染HEK293后对CD2AP细胞内分布的影响。方法:PCR扩增NPHS2cDNA全长,通过T/A克隆连接至pGEMT-easy载体,应用XhoⅠ和BamHⅠ双酶切后,再将NPHS2全长亚克隆入pEGFP-C2。将构建好的荧光表达载体pEGFP-NPHS2转染HEK293细胞,通过免疫荧光的方法观察转染前后CD2AP细胞内的分布情况。结果:①成功构建pEGFP-NPHS2荧光表达载体;②Podocin表达载体转染HEK293细胞后,CD2AP由核周转为弥散的细胞质分布。结论:Podocin转染HEK293细胞可使CD2AP由核周转为弥散的细胞质分布。 Objective：To construct the podocin immunofluorence expression vector and observe its effects of transfection on CD2AP distribution in HEK293 cells. Methods：The pGEMT-easy vector containing the full-length cDNA encoding the human podocin was cloned and digested with BamHI and XhoI. The digested full-length podocin was subcloned into pEGFP-C2. The constructed plasmids were transfected into HEK293 cells,and its effect on CD2AP distribution were observed by immunofluorescence. The interaction between podocin and CD2AP was detected by immunoprecipitation. Results： （1）The pEGFP-NPHS2 expression vector was successfully constructed and podocin exclusively located on HEK293 cell membrane. （2） After podocin transfection, CD2AP redistributed from perinucleus to cytoplasm in HEK293. Conclusion：Podocin can recruit CD2AP to redistribute from perinucleus to cytoplasm in HEK293 cells.
作者 沙玉根 黄松明 张爱华 赵非 陈荣华
机构地区 南京医科大学附属南京市儿童医院肾脏科 南京医科大学儿科研究所
出处 《南京医科大学学报（自然科学版）》 CAS CSCD 北大核心 2007年第6期527-529,533,F0002,共5页 Journal of Nanjing Medical University(Natural Sciences)
基金 国家自然科学基金资助项目(30100081) 南京市卫生局资助项目(ZKM06043)
关键词 PODOCIN 表达载体 CD2AP podocin expression vector CD2AP
分类号 Q74 [生物学—分子生物学] R329.2 [医药卫生—人体解剖和组织胚胎学]
  • 相关文献

参考文献15

  • 1Bounte N,Gribouval O,Roselli S,et al.NPHS2,encoding the glomerular protein podocin,is mutated in autosomal recessive steroid-resistant nephrotic syndrome[J].Nat Genet,2000,24(4):349-354
  • 2Franceschini N,North KE,Kopp JB,et al.NPHS2 gene,nephrotic syndrome and focal segmental glomerulosclerosis:a HuGE review[J].Genet Med,2006,8(2):63-75
  • 3Mao J,Zhang Y,Du L,et al.NPHS1 and NPHS2 gene mutations in Chinese children with sporadic nephrotic syndrome[J].Pediatr Nephrol,2006,21(11):1666-1675
  • 4Hosoyamada M,Yan K,Nishibori Y,et al.Nephrin and podocin expression around the onset of puromycin aminonucleoside nephrosis[J].J Pharmacol Sci,2005,97(2):234-241
  • 5Luimula P,Sandstrom N,Novikov D,et al.Podocyteassociated molecules in puromycin aminonucleoside nephrosis of the rat[J].Lab Invest,2002,82(6):713-718
  • 6Schwarz K,Simons M,Reiser J,et al.Podocin,a raftassociated component of the glomerular slit diaphragm,interacts with CD2AP and nephrin[J].J Clin Invest,2001,108(11):1621-1629
  • 7Lehtonen S,Zhao F,Lehtonen E.CD2-associated protein directly interacts with the actin cytoskeleton[J].Am J Physiol Renal Physiol,2002,283(4):F734-743
  • 8Huber TB,Kottgen M,Schilling B,et al.Interaction with podocin facilates nephrin signaling[J].J Biol Chem,2001,276(45):41543-41546
  • 9Ellin L,Huber TB,Gerke P,et al.NEPH1 defines a novel family of podocin interacting proteins[J].FASEB J,2003,17(1):115-117
  • 10Khoshnoodi J,Sigmundsson K,Ofverstedt LG,et al.Nephrin promotes cell-cell adhesion through homophilie interactions[J].Am J Pathol,2003,163(6):2337-2346

二级参考文献25

  • 1沙玉根,张爱华,陈荣华,徐茵,黄松明,郭梅,费莉.Podocin在大鼠微小病变肾病模型中的表达和分布[J].肾脏病与透析肾移植杂志,2004,13(3):211-214. 被引量：10
  • 2Kestila M, Lenkkeri U, Mannikko M, et al. Positionally cloned gene for a novel glomerular protein nephrin is mutated in congenital nephrotic syndrome[J]. Mol Cell, 1998,1(4): 575-582.
  • 3Shih NY, Li J, Karpitskii V, et al. Congenital nephrotic syndrome in mice lacking CD2-associated protein[J] .Science. 1999. 286(5438): 312-315.
  • 4Khoshnoodi J, Tryggvason K. Unraveling the molecular make-up of the glomerular podocyte slit diaphragm[J] .Exp Nephrol, 2001, 9(6): 355 - 359.
  • 5Boute N, Gfibouval O, Roselli S, et aL NPHS2, encoding the glomerular protein podocin, is mutated in autosomal recessive steroid-resistant nephrotic syndrome[J] . Nat Genet, 2000, 24(4): 349-354.
  • 6Kaplan JM, Kim SH, North KN, et al. Mutations in ACTN4, encoding alpha-actinin-4, cause familial focal segmental glomerulosclerosis[J] . Nat Genet, 2000, 24(3): 251-256.
  • 7Dustin ML, Olszowy MW, Holdorf AD, et al. A novel adaptor protein orchestrates receptor patterningand cytoskeletal polarity in T-cell contacts[J] . Cell, 1998, 94(5): 667-677.
  • 8Palmen T, Lehtonen S, Ora A, et al. Interaction of endogenous nephrin and CD2-assoeiated protein in mouse epithelial M-1 cell line[J]. J Am Soc Nephrol, 2002, 13(7): 1766- 1772.
  • 9Schwarz K, Simons M, Reiser J, et al. Podocin, a raft-associated component of the glomerular slit diaphragm,interacts with CD2AP and nephrin[J]. J Clin Invest, 2001,108(11): 1621-1629.
  • 10Kestila M, Lenkkeri U, Mannikko M, et al. Positionally cloned gene for a novel glomerular protein nephrin is mutated in congenital nephrotic syndrome [J]. Mol Cell,1998,1 (4) :575-582.

共引文献15

同被引文献14

  • 1赵非,黄松明,张爱华,费莉,郭梅,陈荣华.CD2AP、F-actin在肾病大鼠中的表达及意义[J].中国病理生理杂志,2006,22(1):182-186. 被引量：7
  • 2Bertram JF,Messina A,Ryan GB. In vitro effects of puromycin aminonucleoside on the uhrastructure of rat glomerular podocytes[J]. Cell Tissue Res, 1990,260(3): 555-563
  • 3Kawachi H, Miyauchi N, Suzuki K,et al. Role of podocyte slit diaphragm as a filtration barrier [J]. Nephrology (Carlton), 2006, 11 (4) :274-281
  • 4Shaw AS,Miner JH. CD2-associated protein and the kidney[J]. Curr Opin Nephrol Hypertens,2001,10 (1) : 19- 22
  • 5Akhtar M,Al Mana H. Molecular basis of proteinuria[J]. Adv Anat Pathol, 2004, 11 (6) : 304-309
  • 6Dustin ML,Olszowy MW,Holdorf AD,et al. A novel adaptor protein orchestrates receptor patterning and cytoskeletal polarity in T-cell contacts [J]. Cell, 1998,94: 667-677
  • 7Shih NY, Lin J, Karpitskiiv V,et al. Congential nephrotic syndrome in mice lacking CD2-associated protein [J]. Science, 1999,286:312-315
  • 8Tossidou I,Kardinal C,Peters I,et al. CD2AP/CIN85 balance determines receptor tyrosine kinase signaling response in podocytes [J]. J Biol Chem, 2007,282(10): 7457-7464
  • 9Khoshnoodi J,Tryggvason K. Unraveling the molecular make-up of the glomerular podocyte slit diaphragm [J]. Exp Nephrol, 2001,9(6) : 355-359
  • 10Shaw AS. T-cell activation and immunologic synapse [J]. Immunol Res, 2005,32( 1-3 ) : 247-252

引证文献1

二级引证文献1

南京医科大学学报（自然科学版）
2007年 第6期

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