Rapid communication: nucleotide sequence of the ovine lipoprotein lipase cDNA.

Abstract

Name of the Sequence. Ovine lipoprotein lipase cDNA. Genus and Species. Ovis aries. Origin of Clones. Because almost the whole 3′-untranslated region (3′UTR) was found to be lacking from the ovine lipoprotein lipase (LPL) cDNA sequence (1,656 bp including the 5′UTR, the coding sequence, and the first 44 bp of the 3′UTR) published by Edwards et al. (1993), we have undertaken to characterize this region, with the aim to provide a full-length LPL cDNA. First, a 779-bp LPL 3′UTR cDNA was obtained from ovine adipose tissue by rapid amplification of cDNA end (3′RACE) using the following primers: forward 5′-GTATAGTGGCCAAATAGCACA-3′, reverse 5′-TCAAGCTTCTGCAGGATCCTTTTTTTTT TTTTTTTT-3′. The reverse primer was also used to prime reverse transcription. The RACE products were cloned into SmaI-digested and dephosphorylated pGEM-4Z. The LPL recombinant plasmids were screened by PCR performed directly on colonies. Second, the sequence of the fragment lacking between the previous 779-bp fragment and the fragment sequenced by Edwards et al. (1993) was amplified by PCR with the forward 5′-CAAGTCTCTGAATAGAAAGTCT-3′ and reverse 5′-GATTTCCAGTAATAGCCTCTG-3′ primers. Sequencing reactions were carried out on both strands, using an ABI 373A automated DNA sequencer and the accompanying software (PE Applied Biosystems, Courtaboeuf, France).