2004-02-26T00:00
AUTHORSWU, XIAOBING
ABSTRACTThis invention relates to a recombinant adeno-associated virus vector containing RNAi (RNA interference) nucleotide, by which the gene of RNAi nucleotide can be transferred and copied. Thus RNAi nuclenotide can have a biological function. Especially, in this invention we construct a short hairpin-loop expressing vector inhibiting expression of the leuciferinase in BHK-21 cells. The steps including: using human genomic DNA as template, designing primers, U6snRNA promotor is produced by PCR. This promotor is ligated with reverse repeats of the 21bp leuciferinase sequence having 9bp interval and AAV plasmid psNAV in order to construct leuciferinase RNAi expressing plasmid pSNAV/U6/Luc which is cotransfected into BHK-21 cells with plasmid pMAMneoLuc. And pSNAV/U6/Luc and pMAMneoLuc are separately transfected into leuciferinase cells, then the inhibiting effect on leuciferinase expressing, is detected. The result exhibit that the ratio of inhibition is 50 % in the former and 70 % in the latter which confirms that the leuciferinase containing short hairpin-loop can successfully inhibit leuciferinase expressing in BHK-21 cells. More... »
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