ABSTRACT
AIM: To clone a novel mouse GABAA-receptor-associated protein like 2 (Gabarapl2) gene, and to analysis its primary function. METHODS: With the aid of computer, the human GABARAPL2 cDNA was used as information probe to search mouse EST database of GenBank for mouse homolog. A series of overlapping EST were found and assembled into an EST contig using Genetics Computer Group (GCG) ASSEMBLY program. The existence of the gene was then identified by experiment. Northern blotting was performed to hybridize [alpha-32P]dATP labeled probe with mRNA of 11 different mouse tissues that had been transferred to the nylon membrane. RESULTS: The novel gene was deposited in GenBank under Accession No AF190644. Its cDNA contained an intact open reading frame and a canonical polyadenylation signal AATAAA followed by polyA. The deduced protein was completely identical to that of human GABARAPL2, and was termed Gabarapl2 by Mouse Gene Nomenclature Committee. The putative protein of Gabarapl2 has a calculated molecular weight of 13 700 and an isoelectric point of 8.56. It was also predicted to contain two protein kinase C phosphorylation sites and one tyrosine kinase phosphorylation site. Northern hybridization showed that Gabarapl2 was expressed as a single 1.35 kb transcript, with high levels in brain, thymus, lung, heart, kidney, and liver, and low in pancreas, testis, small intestine, colon, and stomach. CONCLUSION: A novel mouse Gabarapl2 gene was cloned and identified.
Subject(s)
Receptors, GABA/genetics , Adaptor Proteins, Signal Transducing , Amino Acid Sequence , Animals , Apoptosis Regulatory Proteins , Base Sequence , Brain/metabolism , Cloning, Molecular , DNA, Complementary/analysis , DNA, Complementary/genetics , Humans , Mice , Microtubule-Associated Proteins/genetics , Molecular Sequence Data , Receptors, GABA/analysis , Receptors, GABA-A , Reverse Transcriptase Polymerase Chain Reaction , Thymus Gland/metabolismABSTRACT
AIM: To further explore the etiological mechanism of Wilson's disease (WD) by comparing the changes of biliary trace elements and its clinical phenotype. METHODS: WD patients with different types and conditions (n = 20), non-WD patients with chronic liver damage (n = 22), and healthy volunteers (n = 10; used as controls) were studied. Biliary samples were taken by duodenal drainage. Atom absorption spectrophotometer was used to assay the copper and zinc content of each sample. RESULTS: In WD, the copper content and copper/zinc ratio of biliary juice were evidently lower than those of non-WD patients with chronic liver damage and of healthy controls (F = 14.76, 25.4; 14.92, 26.2 respectively; P < 0.01), while the biliary zinc level had no significant difference from the two non-WD control groups (P > 0.05). There were significant differences in biliary copper excretion among patients with different types and conditions (F = 3.75, P < 0.05; F = 6.20, P < 0.01). CONCLUSION: Copper excretion by liver and the biliary system decreases obviously in WD, which plays a key role in the phenotypic copper retention, and the biliary copper retention is closely related with the severity of hepatic injury and illness.
