Cloning and functional characterization of the ovine Hormone Sensitive Lipase (HSL) full-length cDNAs: An integrated approach

Cloning and functional characterization of the 5′ regulatory region of ovine Hormone Sensitive Lipase (HSL) gene

“…Using the ovHSL full-length cDNA (Lampidonis et al, 2008), we succeeded in isolating the 5′ regulatory region of the gene by adopting the relatively laborious approach of genome walking. Gene-specific primers were synthesized via the utilization of suitable software programs for primer design, according to the previously reported ovHSL cDNA sequence (Table 1).…”

“…The generation of linearly assembled genomic DNA fragments (contigs) resulted in a 2.744 kb clone. Alignment of the isolated genomic segment with the previously characterized ovHSL cDNA clones (Lampidonis et al, 2008) revealed the presence of a 1.448 kb large intron interposing the 5′-untranslated region (5′-UTR) (Figs. 1-3).…”

“…A polymerization step at 67°C for 7 min was finally added after the completion of 25 cycles. For the first genome walking step, genespecific primers were designed according to the previously characterized 5′-end nucleotide sequence of the ovHSL cDNA (GenBank accession number: DQ647325; Lampidonis et al, 2008). Subsequently, the second genome walking step was attained through the utilization of ovHSL gene-specific primers that were appropriately synthesized according to the 5′-end nucleotide sequence of the first genome walking product.…”

The resurgence of Hormone-Sensitive Lipase (HSL) in mammalian lipolysis

“…Gene structure As HSL has been extensively studied in human and other mammalian species, complete and annotated gene sequences have been already deciphered for human (Holm et al, 1988a;Holst et al, 1996;Langin et al, 1993), mouse (Li et al, 1994;Sztrolovics et al, 1997), rat (Fredrikson et al, 1981;Osterlund et al, 1996), pig (Harbitz et al, 1999) and bovine (Cordle et al, 1986;Garton et al, 1988) species. Ovine HSL (ovHSL) has been analytically studied at enzymatic level in adipose tissue (Bauchart, 1993;Chilliard et al, 2000;Khachadurian et al, 1966;Rogdakis et al, 1997;Vernon, 1980;Vernon and Finley, 1985), whereas an integrated approach at a molecular level regarding the cloning and isolation protocols of the ovHSL full-length cDNAs has been recently performed by Lampidonis et al (2008a). Holm et al (1988a) have mapped the human HSL (hHSL) gene onto chromosome 19 (42,905,659 -42,931,578), whereas, as it has been demonstrated by Warden et al (1993), the mouse homologue is localized onto chromosome 7 (26,164,546 -26,181,006).…”

Integrative computational approach for genome-based study of microbial lipid-degrading enzymes

“…In addition, 3D structural analysis methods have been applied to lipid-degrading enzymes in various organisms, including humans and animals (Fendri et al 2006;Lampidonis et al 2008;Cornaciu et al 2011;Ataya 2012;Bouchaâla et al 2015;Ganesan et al 2015;Mahankali et al 2015). Specifically, structural studies of lipid-degrading enzymes in microbes have been reported (Vasel et al 1993;Gaskin et al 2001;Mandrich et al 2004;Jan et al 2015).…”