Abstract
ATP-citrate lyase (ACL), one of the lipogenic enzymes, catalyses the formation of acetyl-coenzyme A (CoA) involved in the synthesis of fatty acid and cholesterol. In pig, very little is known about the ACL gene. In this work, the mRNA differential display technique was used to analyse the differences in gene expression between Meishan and Large White pigs and the F1 hybrids of both direct and reciprocal crosses. Our results show that among the differentially expressed genes ACL is up-regulated in the backfat of the F1 hybrids. After cloning and analysing the fulllength cDNA and the 870 bp 5'-flanking sequence of the porcine ACL gene, a C/T mutation at position -97 bp upstream of the transcription site was detected. Luciferase activity detection showed that this mutation changed the transcriptional activity. In F1 hybrids, the heterozygous genotype CT was more frequent than the homozygous genotypes CC and TT. Real-time PCR analysis showed that in Meishan pigs, ACL mRNA expression was more abundant in individuals with genotype CT than in those with genotype CC or TT or in Large White pigs. These results indicate that the C/T mutation affects ACL mRNA expression, probably via the activator protein 2.
Highlights
Significant phenotypic differences exist between Chinese indigenous Meishan pigs and western commercial Large White pigs
Semi-quantitative RT-PCR analysis showed that the expression level of EST39 in backfat was higher in the F1 hybrids than in their parents (Fig. 1B)
Sterol regulatory element-binding proteins (SREBPs), especially SREBP-1, regulate ATP-citrate lyase (ACL) enzyme activity at the transcriptional level, whereas NF-Y binding is a prerequisite for activating the ACL promoter [14]
Summary
Significant phenotypic differences exist between Chinese indigenous Meishan pigs and western commercial Large White pigs. The latter present higher growth rate, carcass lean meat percentage and feed to body weight conversion ratio, Article published by EDP Sciences Z.-Q. In our laboratory, using suppression subtractive hybridization (SSH) and mRNA differential display, we have detected significant differences in mRNA quantities and expression patterns for several genes between porcine F1 hybrids and their parents [12,17,19]. Cloning and characterizing genes that are differentially expressed between hybrids and their parents should provide further insights into the genetic basis of phenotypic differences. We show that, in backfat, the gene ATP-citrate lyase (ACL) is differentially expressed between Meishan · Large White and Large White · Meishan F1 hybrids and their parents
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