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Control of Circadian Rhythms and Photoperiodic Flowering by the Arabidopsis GIGANTEA Gene
Deok Hoon Park, David E. Somers, Yang Suk Kim, Yoon Hi Choy, Hee Kyun Lim, Moon Soo Soh, Hyo Jung Kim, Steve A. Kay, and Hong Gil Nam

Supplementary Material

Supplemental Figure 1. Positional cloning of the GI gene. (A) A bacterial artificial chromosome (BAC) contig map around the GI locus and the structure of the GI gene. The positions of the GI and TH1 genes are noted. The GI gene spans the two BAC clones, T23L3 and T22J18. The positions of the exons (black bars) and the mutations sites in gi-1 (open triangle) and gi-2 (solid triangle) are indicated. (B) Identification of restriction fragment length polymorphism (RFLP) pattern in the GI mutants probed with the BAC clone T22J18. An RFLP between wild type (Col-O) and gi-1 was found with the restriction enzyme Stu I and is noted with arrows. The 7.1- and 1.1-kb fragments (isolated from the T22J18 clone) present in the BAC clone and wild-type genomic DNA were absent in the gi-1 mutant and were replaced by a single 8.2-kb fragment. (C) Identification of RFLP fragments. When used as probes, both the 7.1- (left) and 1.1-kb (right) fragments isolated from the T22J18 BAC clone hybridized to the 8.2-kb fragment in gi-1. (D) The predicted amino acid sequences of the GI gene. The predicted transmembrane domains (TMpred, http://www.ch.embnet.org/software/TMPRED_form.html) are underlined. The deletion sites in gi-1 (open triangle) and gi-2 (solid triangle) are noted. Single-letter abbreviations for the amino acid residues are as follows: A, Ala; C, Cys; D, Asp; E, Glu; F, Phe; G, Gly; H, His; I, Ile; K, Lys; L, Leu; M, Met; N, Asn; P, Pro; Q, Gln; R, Arg; S, Ser; T, Thr; V, Val; W, Trp; and Y, Tyr. (E) Structures of the gi-1 and gi-2 mutant alleles. (F) Genomic DNA blot analysis of the GI gene with the full-length cDNA clone as a probe. (G) RNA blot analysis of the GI gene with the full-length cDNA clone as a probe. Ten micrograms of total cellular RNA isolated from root, rosette leaf, stem, cauline leaf, and whole floral organs were used.

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