Molecular cloning and expression of a novel human cDNA related to the diazepam binding inhibitor.
In order to isolate the unidentified autoantigens in autoimmune diabetes, a human pancreatic islet cDNA library was constructed and screened with the sera from the diabetic patients. From the library screening, one clone (DRS-1) that strongly reacted with the sera was isolated. Subsequent sequence analysis revealed that the clone was a novel cDNA related to the diazepam binding inhibitor. DRS-1 was expressed in most tissues including liver, lung, tonsil, and thymus, in addition to pancreatic islets. DRS-1 was in vitro translated and the recombinant DRS-1 protein was expressed in Escherichia coli and purified. The size of the in vitro translated or bacterially expressed DRS-1 protein was in agreement with the conceptually translated polypeptide of DRS-1 cDNA. Further studies are required to test whether or not DRS-1 is a new autoantigen in autoimmune diabetes.
Suk
K
K
Clinical Research Center, Samsung Biomedical Research Institute, Seoul, South Korea.
Kim
Y H
YH
Hwang
D Y
DY
Ihm
S H
SH
Yoo
H J
HJ
Lee
M S
MS
D000595:Amino Acid Sequence; D001324:Autoantigens / Q000235:genetics; D001483:Base Sequence; D019748:Carbon-Carbon Double Bond Isomerases; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D003922:Diabetes Mellitus, Type 1 / Q000235:genetics; D064006:Dodecenoyl-CoA Isomerase; D015723:Gene Library; D006801:Humans; D007515:Islets of Langerhans / Q000378:metabolism; D007535:Isomerases; D008969:Molecular Sequence Data; D012313:RNA / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics*; D016415:Sequence Alignment
D001324:Autoantigens; D018076:DNA, Complementary; D011994:Recombinant Proteins; D012313:RNA; D007535:Isomerases; D019748:Carbon-Carbon Double Bond Isomerases; D064006:Dodecenoyl-CoA Isomerase; C119738:ECI2 protein, human