Abstract
A study was made of the regulatory regionof the gene responsible for the expressionof phycoerythrin in the red alga Gracilaria lemaneiformis.Rapid AmplifiedcDNA Ends Polymerase Chain Reaction(RACE-PCR) was used to obtain the 5'untranslated sequence of phycoerythrinoperon of the alga. The transcriptionalinitiation site starting with A was foundat 80 bp ahead of the AUG codon of thestructural gene. A 54 bp fragment with twoor three gaps inside was determined to be66.7% $sim; 85.2% similar to those of other redalgae.In vitroPCR cloning technique wasused to clone the far upstreamuntranscribed sequence. There was nosignificant similarity of untranscribedsequence to those known for other redalgae. Neither a canonical TATA nor CAATbox was found in this region. As predictedby PLACE, a software suitable for plantpromoter analysis, there were two specialfunctional area, one around nucleotide 65,the other 328, responsible for lightregulated and /or high level of geneexpression. The untranscribed region ofphycoerythrin operon was subcloned into apromoter report vector, pEGFP, andtransferred intoE. coliand SynechocystisPCC6803, respectively.Strong green fluorescence was observed inelectroporatedSynechocystisPCC 6803cells 24hrs after the operating, but not inE. coli, suggesting that the promoteris trans-activated.
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