cDNA sequence and tissues expression analysis of lipoprotein lipase from common carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis> Var. Jian) |
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Authors: | Han-liang Cheng Si-ping Sun Yong-xing Peng Xiao-yun Shi Xin Shen Xue-ping Meng Zhi-guo Dong |
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Institution: | (1) College of Marine Science, Huaihai Institute of Technology, No. 59 Cangwu Road, 222005 Lianyungang, People’s Republic of China |
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Abstract: | A full-length cDNA coding lipoprotein lipase (LPL) was cloned from liver of adult common carp (Cyprinus carpio Var. Jian) by RT-PCR and rapid amplification of cDNA ends (RACE) approaches. The cDNA obtained was 2,411 bp long with a 1,524 bp
open reading frame (ORF) encoding 507 amino acids. This amino acid sequence contains two structural regions: N-terminus (24–354
residues) and C-terminus (355–507 residues). Before N-terminus, 1–23 residues is signal peptide, 6–23 residues is transmembrance
helix. At N-terminus, some conversed functional sites were found, including two N-linked glycosylation sites Asn41 and Asn88; one catalytic triad Ser174, Asp198 and His283; one conserved heparin-binding site Arg321 to Arg324 (RKNR); eight cysteines residues Cys69 and Cys82, Cys258 and Cys281, Cys306 and Cys325, Cys317 and Cys320 which are involved in four disulfide bridges; one polypeptide “lid” that participates in substrate specificity. At C-terminus,
Asn401 is another N-linked glycosylation site, and Trp434 and Trp435 (WW) is lipid-binding site. The amino acid sequence has a high similarity, and shows similar structural features to LPL of
other species. Tissue distribution of LPL mRNA in liver, head kidney, mesenteric adipose tissue, heart and white muscle of common carp was analyzed by semi-quantitative
RT-PCR method using β-actin gene as internal control. The result showed that the expressions of LPL mRNA were detected in all examined tissues of common carp. The expression levels of LPL in the mesenteric adipose tissue was highest among these tissues, following in liver and head kidney, and the lowest expression
was found in heart and white muscle. |
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