Identification of genes regulated by glucose in a pancreatic beta-cell line by a new method for subtraction of mRNA.
Various genes are differentially expressed in cells during cell differentiation, development, aging, and in pathological conditions. To identify and isolate the genes that are specifically and differentially expressed in cells, we established a ligation-mediated polymerase chain reaction (PCR)-based method for cDNA subtraction. As this method is PCR-based, when target genes are expressed at high levels relative to the driver (a control pool for subtraction), even a small amount of target genes can be amplified. By this newly developed PCR-based subtraction method, a set of genes regulated by glucose were identified in a mouse insulinoma cell line. This PCR-based and non-radioactive subtraction method will be a powerful tool for identification of novel genes, specifically and differentially expressed in cells.
Yamato
E
E
Department of Geriatric Medicine, Osaka University Medical School, Japan.
Ikegami
H
H
Miyazaki
J I
JI
Ogihara
T
T
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D005786:Gene Expression Regulation / Q000235:genetics*; D015723:Gene Library; D005947:Glucose / Q000235:genetics / Q000502:physiology*; D007515:Islets of Langerhans / Q000166:cytology / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009928:Organ Specificity; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics
D017931:DNA Primers; D018076:DNA, Complementary; D012333:RNA, Messenger; D005947:Glucose