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The Expression of Anti-bcl-2 Ribozyme Gene in HL-60 Cells
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Abstract:
A hammerhead RZ DNA was designed and synthesized, which can specifically cleave the bcl-2 mRNA. After demonstration of right sequences by sequencing and cleavage activity of RZ by in vitro cleaving experiment, The RZ DNA was recombinated into the pDOR - neo vector to form the recombinant pDOR - RZ. Using lipofectin - mediated DNA transfectionpDOR-RZ was successfully introduced into HL - 60 cells. The RZ expression was observed by Southern, RNA dot blot hybridization and flow cytometry (FCM) . The results demonstrated that (a) the RZ was expressed in 72 hours after transfection; (b) the synthesis of Bel - 2 protein was inhibited by the expression of RZ; (c) apoptotic peak appeared in FCM.
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