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Background: Ghrelin is a multifunctional peptide hormone expressed in a range of normal tissues and pathologies. It has been reported that the human ghrelin gene consists of five exons which span 5 kb of genomic DNA on chromosome 3 and includes a 20 bp non-coding first exon (20 bp exon 0). The availability of bioinformatic tools enabling comparative analysis and the finalisation of the human genome prompted us to re-examine the genomic structure of the ghrelin locus., Results: We have demonstrated the presence of an additional novel exon (exon -1) and 5' extensions to exon 0 and 1 using comparative in silico analysis and have demonstrated their existence experimentally using RT-PCR and 5' RACE. A revised exon-intron structure demonstrates that the human ghrelin gene spans 7.2 kb and consists of six rather than five exons. Several ghrelin gene-derived splice forms were detected in a range of human tissues and cell lines. We have demonstrated ghrelin gene-derived mRNA transcripts that do not code for ghrelin, but instead may encode the C-terminal region of full-length preproghrelin (C-ghrelin, which contains the coding region for obestatin) and a transcript encoding obestatin-only. Splice variants that differed in their 5' untranslated regions were also found, suggesting a role of these regions in the post-transcriptional regulation of preproghrelin translation. Finally, several natural antisense transcripts, termed ghrelinOS (ghrelin opposite strand) transcripts, were demonstrated via orientation-specific RT-PCR, 5' RACE and in silico analysis of ESTs and cloned amplicons., Conclusion: The sense and antisense alternative transcripts demonstrated in this study may function as non-coding regulatory RNA, or code for novel protein isoforms. This is the first demonstration of putative obestatin and C-ghrelin specific transcripts and these findings suggest that these ghrelin gene-derived peptides may also be produced independently of preproghrelin. This study reveals several novel aspects of the ghrelin gene and suggests that the ghrelin locus is far more complex than previously recognised.

Microalgae already serve as a major natural source of valuable macromolecules including carotenoids, long-chain polyunsaturated fatty acids and phycocolloids. As photoautotrophs, their simple growth requirements make these primitive plants potentially attractive bioreactor systems for the production of high-value heterologous proteins. The difficulty of producing stable transformants has meant that the field of transgenic microalgae is still in its infancy. Nonetheless, several species can now be routinely transformed and algal biotechnology companies have begun to explore the possibilities of synthesizing recombinant therapeutic proteins in microalgae and the engineering of metabolic pathways to produce increased levels of desirable compounds. In this review, we compare the current commercially viable bioreactor systems, outline recent progress in microalgal biotechnology and transformation, and discuss the potential of microalgae as bioreactors for the production of heterologous proteins.

We report the finding of two non-allelic genes encoding tyrosine hydroxylase (TH) from the diploid teleost barramundi Lates calcarifer. Barramundi TH1 is the homologue of the higher vertebrate TH genes and encodes a protein of 489 amino acids that shares 90% sequence identity to the THs of other teleost species. A second non-allelic tyrosine hydroxylase gene (TH2) encodes a protein of 472 amino acids and shares 62% identity with TH1 and the vertebrate THs. TH1 mRNA is found in the brain and kidney of barramundi while TH2 mRNA is found only in brain. The TH2 gene is also present in the genomes of the pufferfish Takifugu and zebrafish Danio. Estimates of the rates of nucleotide substitution suggest the teleost TH2 genes are selectively constrained although not to the degree seen in the TH1 genes. Differential regulation of the two TH genes is, however, indicated by differences in transcript distribution, the nature of the Ca2+-responsive elements found in the proximal promoter region and the lack of recognised phosphorylation sites in TH2. Preservation of two apparently functional TH genes in phylogenetically distant teleost species is consistent with the notion of partitioning of function between duplicate genes.