The Construction Of Transgenic Zebrafish Overexpressing Human BAFF And The Detection Of Immune-related Genes Expression
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    Abstract:

    Objective:to study the pathogenesis of autoimmune disease, transgenic zebraifsh overexpressing human BAFF is constructed . Method:855bp BAFF cDNA was cloned by RT-PCR from human lymphoma cell line to construct the BAFF overexpression recombinant plasmid Tol2-hBAFF. The protein expression in Hela cell-line was detected by western blot after invitro transfection. The zebrafish embryo was injected with recombinant BAFF plasmids and screened by GFP tracing. The GFP positive embryos were selected to analyze the early immune-related genes expression by qPCR. Result: BAFF-GFP fusion protein was successfully expressed, which proved an effective method to produce the transgene zebrafish for future study. Besides, qPCR results demonstrated that TCRAC signficantly high expressed in 1dpf transgenic embryos while the Ikaros gene was down-regulated. It suggests that BAFF overexpression in zebrafish embryo may reslult in immuno-related gene premature expression in early lymphatic system. All this results proved that transgenic zebrafish can be obtained by microinjection of the Tol2-hBAFF recombinant plasmids in zebrafish embryo, which supply an effective method to explore the human SLE disease and its antagonist on animal models.

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History
  • Received:April 20,2013
  • Revised:June 05,2013
  • Adopted:June 15,2013
  • Online: October 18,2013
  • Published: