Identification of SKIP homologs in soybean. (A) Predicted domains and motifs of soybean SKIP homolog, GmGBP1. (B) Multiple alignment of the predicted amino acid sequences in peptide signature of SKIP homologs from soybean (GmGBP1, DQ112540), Arabidopsis (AtSKIP, O80653), rice (OsSKIP,), human (HsSKIP, Q13573), Caenorhaabditis elegans (CeSKIP, Q22836), Drosophila (DmBX42, P39736), Saccharomyces cerevisiae (ScPRP45, Q09882). (C) Detection for self-transactivation of GmGBP1. The deletion mutants (shown on right), GmGBP1, GmGBP1a (amino acids 1–189), GmGBP1b (amino acids 190–356) and GmGBP1c (amino acids 357–612), were constructed by PCR. Self-transactivation, as illustrated by the color reaction in the 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside(X-Gal) assay, occurred with 1h for pBD-GmGBP1 and pBD-GmGBP1c constructs. Overnight incubation failed to produce a self-activation reaction with the pBD-GmGBP1a and pBD-GmGBP1b constructs. And the transformants were streaked on SD agar plates lacking His, Trp and incubated at 30°C for 72 h. The pBD-GmGBP1 and pBD-GmGBP1c constructs could grow normally but no growth for the pBD-GmGBP1a and pBD-GmGBP1b constructs. (D) Phylogenetic relationships of SKIP homologs in plant constructed using the neighbor-joining method with the program MEGA 5.05.
Zhang et al. BMC Plant Biology 2013 13:21 doi:10.1186/1471-2229-13-21