中国水产科学Issue(5)：950-957,8.DOI:10.3724/SP.J.1118.2013.00950
虾夷马粪海胆溶菌酶基因全长cDNA的克隆与表达分析
Cloning and expression analysis of a sea urchin(Strongylocentrotus intermedius) lysozyme gene
摘要
Abstract
A full-length cDNA coding lysozyme (LYZ) was cloned from sea urchin Strongylocentrotus intermedius by reverse transcriptase-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends(RACE) methods. The full-length cDNA of LYZ was 912 by with a 480 by open reading frame(ORF) encoding 159 amino acids, which included a signal peptide of 20 amino acids at the N-terminus and a mature peptide of 139 amino acids. The deduced amino acid sequence had a putative size of 17.69 kD and the theoretical isoelectric point was 7.75. The multiple alignments revealed identity of 91.4% between S. intermedius and S. pu}pu}atus and 59.3% identity between S. intermedius and Apostichopus japonicus in the i-type LYZ amino acid sequence. The i-type lysozyme conserved sequence DVGSLSCGP(Y)Y(F)QIK was detected in the S. intermedius amino acid sequence. These results indicate that the cDNA sequence cloned from S. intermedius is a member of the i-type lysozyme family. Real-time quantitative PCR was carried out to measure LYZ mRNA expression in different tissues and monitor LYZ mRNA expression patterns in the coelomic fluid after a lipopolysaccharide (LPS) challenge. LYZ mRNA expression levels in peristome membrane were significantly higher than those of muscle in the Aristotle's lantern, tube-foot, intestines, coelomic fluid, male gonad, and female gonad (P<0.05). LYZ expression in the coelomic fluid was up-regulated after the LPS challenge and reached its maximum level at 8 h post-stimulation, and decreased gradually thereafter. Thirty-six hours after the LPS challenge, LYZ levels in the coelomic fluid were similar to those of the control.关键词
虾夷马粪海胆/溶菌酶/基因表达/脂多糖((LPs)Key words
Strongylocentrotus intermedius/lysozyme/gene expression/lipopolysaccharide(LPS)分类
农业科技引用本文复制引用
姬南京,杨芸菲,丁君,常亚青..虾夷马粪海胆溶菌酶基因全长cDNA的克隆与表达分析[J].中国水产科学,2013,(5):950-957,8.基金项目
国家自然科学基金资助项目(30972269) （30972269）
国家863计划项目(2012AA 1 OA412) （2012AA 1 OA412）
辽宁省科技计划项目(2007203004) （2007203004）
