Molecular cloning of the human ryudocan promoter

The promoter region of human ryudocan was isolated from a human lambda dash genomic library and cloned into pBluescript. The 5'flanking region contained a classical TATA box and GC rich regions that are commonly found in constitutively expressed genes. Two fusion gene constructs, one of 898 bp and the other of 480 bp of the 5' flanking DNA coupled to the luciferase gene, were transiently expressed in a mouse endothelioma cell line and in human umbilical endothelial cell cultures. Both constructs were capable of driving luciferase expression; the 898 bp construct produced greater levels of luciferase activity than the 480 bp construct in both cell types examined. Analysis of the sequence revealed the presence of several potential sites for nuclear transcription factor binding; the relevance of these sites is presently unknown.