Figure 2

Germline expression of integrated transgenes. (A) P _hsp-16.1 ::venus was co-injected with a positive-selection marker, a negative-selection marker and an injection marker. Ex array-carrying transgenic animals were treated by UV/TMP. F1 animals were cultured under the condition of positive-negative selection. PCR selection with primers to detect the intact P _hsp-16.1 ::venus transgene (PCR#C), determined the integrant lines. qPCR3 indicates the amplified region by quantitative PCR described below. (B) Venus protein was expressed in germ cells by heat shock, suggesting that low-copy transgenes were successfully integrated. tmIs894 adult hermaphrodites are presented. Scale bar = 50 μm. (C) Relative amount of the vps-45 gene determined by quantitative PCR with primers that amplified the 5' region in the vps-45 mini gene (gray bars: qPCR1) and the middle region (black bars: qPCR2; left graph), and relative amounts of the hsp-1.2 promoter region (gray bars: qPCR3; right graph). The amount (normalized to the act-2 gene) is presented as a ratio to the N2 control. Error bars represent SE of three independent experiments.