Structural and functional studies of a family of developmentally regulated, prestalk genes coding for small proteins-5

Copyright information:Taken from "Structural and functional studies of a family of developmentally regulated, prestalk genes coding for small proteins"http://www.biomedcentral.com/1471-2180/8/1BMC Microbiology 2008;8():1-1.Published online 3 Jan 2008PMCID:PMC2257962.for over-expression. Group 1 antibodies were used for Western-Blot. The migration of the 10 and 15 kDa protein markers is shown on the left. B. RNA was obtained from WT and two different RNAi clones that expressed an interfering RNA based on exon 2 of (1 and 6 clones) after 14–18 hours of development on Nitrocellulose filters. Arrows indicate the position of the endogenous mRNA (mRNA) and of the RNAi transcribed from the vector (RNAi construct). Ethidium bromide staining of the gel is shown in the lower panel. C. RNA from WT (AX4) and Group 1 KO strains, harvested at the indicated hours of development (16–18 h) was analyzed by RT-PCR for expression of Group 1 genes. The control band obtained by amplification of a region of the large mitochondrial ribosomal RNA (control: 320 bp long) and that corresponding to Group 1 mRNA (:240 bp) are indicated by arrows at the right. D. RNA was obtained from WT (AX4) and two different Group 2 KO strains (KO12, KO20) after 16 hours of development. Expression of Group 2 genes was analyzed by RT-PCR. The migration of the control band (control: 320 bp) and that corresponding to Group 2 mRNA (: 240 bp) is indicated at the right. E. DNA from WT (AX4) and a Group 1 and Group 2 Double KO strain (Double KO) was analyzed by PCR for the presence of for Group 1 genes. Migration of the amplified fragments for a control gene (control: 600 bp) and Group 1 genes (410 bp) is indicated at the right.