|ZFIN ID: ZDB-IMAGE-140506-25|
bucky ball constructs comprised of exons and introns are spliced correctly. A) Schematic of constructs used to generate bucky ball transgenics with introns and exons of bucky ball. Promoters used were beta-actin and bucky ball. B) Gels showing PCR products amplified from genomic DNA using primers within the 52UTR and 32UTR of the bucky ball gene to obtain versions of buc with the introns intact and either full length or a truncated 32UTR. C) Assay for splicing of buc plasmids including exons and introns. The pBH: β-actin:gbuc construct was injected into single cell embryos along with transposase RNA. At 2dpf (a stage when endogenous buc transcripts are not detected), larvae with the integrated transgene were selected based on their fluorescent hearts and used to generate cDNA to examine the transcripts produced from the buc+introns construct. D) Schematic of spliced buc obtained from pBH: β-actin:gbuc. E) PCR fragments amplified from cDNA of pBH:β-actin:gbuc expressing embryos were as expected for properly spliced buc. Yellow asterisks indicate non-specific products amplified in the absense of buc as determined by sequencing.
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