Molecular cloning and tissue distribution of lipoprotein lipase full-length cDNA from Pengze crucian carp (Carassius auratus var. Pengze)

Abstract

A full-length cDNA coding lipoprotein lipase (LPL) was cloned from liver of adult Pengze crucian carp (Carassius auratus var. Pengze) by RT-PCR and rapid amplification of cDNA ends (RACE) approaches. The cDNA obtained was 1877 bp long with a 1524 bp open reading frame (ORF) encoding 507 amino acids, including a putative signal peptide of 23 amino acids long. The deduced amino acid sequence has a high similarity and shows similar structural features to LPL of other species. The LPL protein has a calculated molecular mass of 57.7 kDa and isolectric point of 7.85. Tissue distribution of LPL mRNA in mesenteric adipose tissue, liver, heart, head kidney and white muscle of adult Pengze crucian carp was analyzed by semi-quantitative RT-PCR method using beta-actin gene as internal control, the result showed that this gene was ubiquitously expressed in all tissues tested with the highest abundance in mesenteric adipose tissue, following in head kidney and liver, and the lowest expression was found in heart and white muscle.