Compact intergenic regions of the pufferfish genome facilitate isolation of gene promoters: characterization of Fugu 3′‐phosphoadenosine 5′‐phosphosulfate synthase 2 (fPapss2) gene promoter function in transgenic Xenopus

FEBS Letters

Browder, Leon¹  Venkatesh, Byrappa²  Brenner, Sydney²  Luchman, Artee¹  Johnston, Jillian¹  Stapleton, Tara¹  Jirik, Frank R.¹ 

[1] Department of Biochemistry and Molecular Biology, University of Calgary, 3330 Hospital Drive N.W., Calgary, AB, Canada T2N 4N1;Institute of Molecular and Cell Biology, 30 Medical Drive, Singapore 117609

关键词: 3′-Phosphoadenosine 5′-phosphosulfate synthase 2;    Enhanced green fluorescent protein;    Gene promoter;    Transgenic;    Intergenic region;    Xenopus laevis;    Fugu rubripes;   

DOI  :  10.1016/S0014-5793(03)01353-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The highly compact nature of the pufferfish (Fugu rubripes) genome renders it a useful tool not only for annotating coding regions within vertebrate genomes, but also for the identification of sequences important to gene regulation. Indeed, owing to this compaction it will be feasible in many instances to initiate analyses using entire intergenic regions when mapping gene promoters; a strategy that is very rarely feasible with the expanded genomes of other species. Stemming from our interest in studying promoters expressed in chondrocytes, we selected for study the intergenic region upstream of Fugu 3′-phosphoadenosine 5′-phosphosulfate synthase 2, fPapss2, a gene required for the normal development of cartilage extracellular matrix. Functional characterization of the entire fPapss2 5′ intergenic region was carried out by monitoring expression of the enhanced green fluorescent protein (EGFP) gene reporter in the developing cartilage of transgenic Xenopus laevis. By evaluating a series of 5′ intergenic region deletions we defined a minimal fPapss2 sequence of ∼300 bp that was essential for EGFP expression in tadpole cartilage. This functional analysis of an entire Fugu intergenic region, combined with the efficiency of Xenopus transgenesis, serves as a model for the rapid characterization of evolutionarily-conserved regulatory regions of other pufferfish genes.

【 授权许可】

Unknown