›› 2008, Vol. 28 ›› Issue (6): 361-366.

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Study on Expression of tau in DJ-1 and DJ-1L166P Transfected NIH3T3 Cells

  

  1. The Key Lab of Laboratory Transgenic Animal ofLiaoNingProvince, Department of Laboratory Animal Science, China Medical University, Shenyang 110001,China
  • Received:2008-09-05 Online:2008-06-30 Published:2008-06-30

Abstract: Objective To explore the relationship between the expression of tau and DJ-i gene in transfected NIH3T3 cell by pEGFP-DJ-1 and pEGFP-DJ-iL166Pplasmid, and it would provide experimental basis to establish DJ-1 and DJ-iL166P transgenic mice and to understand the function of DJ-1 and the pathogenesis of Parkinson’s disease. Methods DJ-1 cDNA sequence were obtained from total RNAof SH-SY5Y cells by RT-PCR. Then one clone of the cDNA was treated with a mutation kit to get a mutation at the site which encodes the 166th amino acids. Recombinant pEGFP-ZV-i and pEGFP-DJ-iL166P plasmids were constructed in a routine way. NIH3T3 cells were transfected into pEGFP-ZV-i,pEGFP-DJ-IL166P, or pEGFP-C3 plasmids, and then the cells were selected with G418. The successfully transfected cells were analyzed by RT-PCR and Western blot to detect the expression of tau gene. Results We gained 6,2, and 9 clones of pEGFP-DJ-i, pEGFP-DJ-IL166P, and pEGFP-C3 transfected cells. The results of RT-PCR and Western blot show that: the expression of tau in pEGFP-DJ-i transfected cells is lower than in pEGFP-C3 transfected cells, and the expression of tau in pEGFP-DJ-IL166P, transfected cells is higher than in pEGFP-C3 transfected cells. The experimental result analyzed by statistics method is significant (P<0.05). Conclusion DJ-1 gene down-regulates the expression of tau on both transcription and translation levels, and DJ-IL166P, up-regulats the expression of tau. The expression of tau is associated with DJ-i gene in NIH3T3 cell.

Key words: DJ-1, tau, Parkinson’s disease