Porcine UCHL1: genomic organization, chromosome localization and expression analysis

摘要

The human UCHL1 gene encodes the ubiquitin C-terminal hydrolase UCHL1, which comprises more than 2% of total brain protein. UCHL1 is a component of the ubiquitin-proteasome system, which degrades overexpressed and damaged proteins. Mutations in the UCHL1 gene are associated with susceptibility to and protection from Parkinson's disease. Here we report cloning, characterization, expression analysis and mapping of porcine UCHL1. The UCHL1 cDNA was amplified by reverse transcriptase polymerase chain reaction (RT-PCR) using oligonucleotide primers derived from in silico sequences. The porcine cDNA codes for a protein of 223 amino acids which shows a very high similarity to human (98%) and to mouse (97%) UCHL1. In addition, the genomic organization of the porcine UCHL1 gene was determined. The porcine UCHL1 gene was mapped to chromosome 8(A1/2p21)-p23. Three SNPs were found in the porcine UCHL1 sequence. Expression analysis by quantitative real time RT-PCR demonstrated that porcine UCHL1 mRNA is differentially expressed in various organs and tissues and similar to its human counterpart. UCHL1 transcript is most abundant in brain tissues and in the spinal cord. The UCHL1 mRNA expression was also investigated in developing porcine embryos. UCHL1 transcript was detected as early as 40 days of gestation. A significant decrease in UCHL1 transcript was detected in basal ganglia from day 60 to day 115 of gestation.