Dear specialists in TDP 43 researches in Drosophila Melanogaster
I actually constructed human TDP 43 in pacu2 vector(uas promoter included) and injected it into Drosophila.
and checked how it expresses.
however, it turns out the transgenic fly i made could not produce TDP 43 proteins even though RNA expression is well characterized. Currently we could not find any possible reason.
below is the construct i designed, please check if there is any mistake.
Kozak-ATG(start)-V5(epitope)-Linker(GGGGS)-TDP43-Linker-2xAU1(epitope)-stop
as i said on title i did not execute codon optimization for human TDP 43 sequences.
and i also skipped first ATG sequence inside human TDP 43 as it exists just behind Kozak sequence.
if anyone see a possible error here please let me know
Thanks.
No, codon optimization shouldn't be necessary.. many TDP-43 constructs used in Drososphila research have not used codon-optimization and they work fine. Perhaps the tags on either end of the construct may have de-stabilized the protein..
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