ISSN:
1432-0428
Keywords:
Key words Random cDNA sequencing
;
MIN6
;
pancreatic islet beta cell.
Source:
Springer Online Journal Archives 1860-2000
Topics:
Medicine
Notes:
Summary To understand the molecular basis of glucose concentration-responsive insulin synthesis and secretion from pancreatic islet beta cells, a group of pancreatic islet beta-cell-related cDNAs was cloned. A pair of cDNA libraries was constructed from a mouse pancreatic islet beta-cell line of MIN6, which was cultured in either high glucose or low glucose media. By applying a random cDNA sequencing approach, 503 and 395 independent species were obtained from a total of 1,011 and 762 clones in the high glucose and low glucose library, respectively. The unknown genes comprised the majority of about 70 % independent clones in both libraries. In Northern blot analysis, 311 (69.4 %) of 448 independent clones showed positive signals within 72 h of autoradiographic exposure. Surprisingly, 150 (48.2 %) out of 311 positive clones showed positive signals to MIN6 cells, but not to NIH/3T3 fibroblasts. The expression level of three unknown clones were glucose-concentration dependent. Combination of a random cDNA sequencing approach and Northern blot analysis is useful to obtain a large number of novel genes and islet beta-cell-related genes. [Diabetologia (1995) 38: 381–386]
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00410274
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