Effect of 5'-flanking sequence deletions on expression of the human insulin gene in transgenic mice

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M Fromont-Racine, D Bucchini, O Madsen, P Desbois, S Linde, C Saulnier, M A Ripoche, J Jami, R Pictet, Jens Høiriis Nielsen

Expression of the human insulin gene was examined in transgenic mouse lines carrying the gene with various lengths of DNA sequences 5' to the transcription start site (+1). Expression of the transgene was demonstrated by 1) the presence of human C-peptide in urine, 2) the presence of specific transcripts in pancreas, but not in other tissues, 3) the specific immunofluorescence staining of pancreatic islets for human C-peptide, and 4) the synthesis and accumulation of human (pro)insulin in isolated islets. Deletions in the injected DNA fragment of sequences upstream from positions -353, -258, and -168 allowed correct initiation of the transcripts and cell specificity of expression, while quantitative expression gradually decreased. Deletion to -58 completely abolished the expression of the gene. The amount of human product that in mice harboring the longest fragment contributes up to 50% of the total insulin does not alter the normal proportion of mice insulins I and II. These results suggest that expression of the human insulin gene in vivo results from the cooperation of several cis-regulatory elements present in the various deleted fragments. With none of the deletions used, expression of the transgene was observed in cell types other than beta-islet cells.
OriginalsprogEngelsk
TidsskriftMolecular endocrinology (Baltimore, Md.)
Vol/bind4
Udgave nummer5
Sider (fra-til)669-77
Antal sider9
ISSN0888-8809
StatusUdgivet - maj 1990

ID: 47974095