Fig. 2

Generation of the tbc1 null allele. a Genomic location of tbc1. The deficiency Df(2 L)Exel6004, which deletes tbc1 and all or part of another 15 genes, is indicated. Small arrows indicate direction of transcription for each of the nearby transcripts. The region removed in the knockout is indicated. tbc1 KO F and R primers were used with pW25–2 or PW25–3, respectively, to confirm the knockout. The thick black line represents the homologous regions used for generating the knockout. The numbers next to and just below the white + insertion indicate exactly which sequences were removed by homologous recombination. b PCR of two tbc1 knockout lines, 178 and 305. Line 178 was used for all the experiments throughout the rest of this work. The negative control for the PCR is one of the lines that is transgenic for the original knockout construct, and maps to a different chromosome. Primers used are those indicated in A. Arrow shows the expected band size for the 5′ end, the arrowhead shows the expected band size for the 3′ end. Asterisks indicate multiple bands also found in the negative control. c In situ stains of tbc1 and lacZ mRNA in heterozygous and homozygous tbc1 knockout siblings. Green arrows indicate lacZ expression from the balancer chromosome; Black arrow indicates the salivary gland. tbc1 mRNA is absent when there is no lacZ mRNA (i.e in homozygous knockout embryos)