National Committee on Marine Sciences (NCMS)
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- Expressed sequence tags (ESTs) from the marine red alga Gracilaria gracilisLluisma, Arturo O.; Ragan, Mark A. (Springer, 1997-06)Expressed sequence tags (ESTs) are partial sequences of cDNAs, and can be used to characterize gene expression in organisms or tissues. We have constructed a 200-sequence EST database from vegetative thalli of Gracilaria gracilis, the first ESTs reported from any alga. This database contains recognizable ESTs corresponding to genes of carbohydrate metabolism (seven), amino acid metabolism (three), photosynthesis (five), nucleic acid synthesis, repair and processing (three), protein synthesis (14), protein degradation (six), cellular maintenance and stress response (three), other identifiable protein-coding genes (13) and 146 sequences for which significant matches were not found in existing sequence databases. We have already used this EST database to recover genes of carbohydrate biosynthesis from G. gracilis.
- Occurrence of closely spaced genes in the nuclear genome of the agarophyte Gracilaria gracilisLluisma, Arturo O.; Ragan, Mark A. (Springer, 1999)Little is known about the structure and organisation of nuclear genomes in red algae. In particular, it is not known whether genes are densely or loosely packed, whether gene order is conserved, whether their genes tend to occur in one or multiple copies and whether their nuclear genes tend to be compact or interrupted by numerous introns. Sequencing of cloned genomic DNA from Gracilaria gracilis has begun to provide provisional answers to some of these questions. Four pairs of closely spaced genes have been found in G. gracilis upon sequencing genomic clones that contain genes for UDPglucose pyrophosphorylase, galactose-1-phosphate uridylyltransferase, the β subunit of tryptophan synthetase, and methionine sulphoxide reductase (a fifth pair of closely spaced genes, encoding polyubiquitin and aconitase, was reported earlier). An open reading frame with significant similarity to another known gene occurs close (< 1.7 kbp) to each of these genes. In two pairs the intergenic region is less than 400 bp in length, and for these the location of the putative polyadenylation signals indicates that the gene transcripts, encoded on opposite strands, have overlapping (hence complementary) 3′ regions. These somewhat unexpected findings begin to establish a basis for genome-level characterisation of red algae.
- Cloning and characterization of a nuclear gene encoding a starch-branching enzyme from the marine red alga Gracilaria gracilisLluisma, A. O.; Ragan, M. A. (Springer, 1998-08-27)The biosynthesis of starch in red algae occurs in the cytosol, in contrast to green plants where it takes place in the plastid. We have cloned a nuclear gene from the red alga Gracilaria gracilis that encodes a homolog of starch-branching enzymes (SBEs); this gene, which is apparently intron-free, was designated as GgSBE1. A potential TATA box, CAAT boxes, and other potential regulatory elements were observed in its 5′ flanking region. The encoded 766-aa peptide shares significant sequence similarity with SBEs from green plants (at least 40%), and with glycogen-branching enzymes (GBEs) from human (46%) and Saccharomyces cerevisiae (45%). Southern-hybridization analysis indicates that the gene is single-copy, although weaker signals suggest that related genes exist in the genome of G. gracilis. Phylogenetic analyses indicate that GgSBE1 groups within the eukaryote branching enzymes (BEs) and not with eubacterial GBEs, suggesting that its gene has not been derived directly from an endosymbiotic cyanobacterium, but instead is ancestrally eukaryotic.