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Cloning and characterization of human ubiquitin binding enzyme 2 cDNA
Guangtao Li1, Hongyan Lü, Yan Zhou
1National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS & PUMC, Chinese National Human Genome Center, Beijing 100005.
Objective:
To clone and identify the gene encoding human ubiquitin binding enzyme 2 and study its expression pattern.
Methods:
According to the sequence of human EST, which is highly homologous to the mouse ubiquitin binding/conjugating enzyme (E2), primers were synthesized to screen the human fetal brain cDNA library. The gene was analyzed by bioinformatics technique and its expression pattern was studied by using multiple-tissue Northern blot.
Results:
Two cDNA clones encoding human ubiquitin conjugating enzyme have been isolated and identified. Both containing the ubiquitin conjugating domain, the 2 cDNA clones are 88% identical in amino acid sequences and splicing isoforms to each other only with an exon excised to form the short sequence. They belong to a highly conserved and widely expressed E2 enzyme family. Northern blot shows that they are expressed exclusively in adult human heart, placenta, and pancreas but no transcripts can be detected in brain, lung, liver, skeletal muscle or kidney.
Conclusions:
The gene encoding human ubiquitin binding enzyme is expressed under temporal control. As a key enzyme in the degradation of proteins, ubiquitin conjugating enzymes play a central role in the expression regulation on the level of post-translation.