Cloning and Analysis of Fhx/P25 Gene Promoter of Bombyx mori Fibroin Protein
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Graphical Abstract
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Abstract
In order to better understand the model of the molecular mechanisms governing spatially and temporally programmed transcription, the Fhx/P25 gene promoter of the fibroin protein from Bombyx mori was cloned and sequenced. An expression vector named pSK-P25-DsRed-PolyA was constructed, in which the reporter gene DsRed was driven by Fhx/P25 promoter. The promoter's activity was then characterized by transient expression assays in BmN cells of B. mori. The results of a sequence analysis showed that the Fhx/P25 promoter possesses the characteristics both of a eukaryotic promoter and a silk gland-specific expression promoter. A conserved TATA box sequence was located at position -28－-32 and had the sequence TATAA. There were three CAAT motifs, of which the two CAAT motifs located at positions -110－-117 and -90－-87 may be active. Secondary structure analysis indicated that the sequence of the promoter region forms a complicated stemloop structure. This may relate to the tissue speciality or the timing and activity of the protein expression. The transient expression product of the red fluorescent gene, driven by the promoter Fhx/P25, can be observed in cultured BmN cells of B. mori.
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