海洋科学2016，Vol.40Issue(3)：32-39,8.DOI:10.11759/hykx20150113001
海带磷酸甘露糖变位酶（PMM）基因的克隆与表达分析
Cloning and expression of phosphomannomutase from Sac-charina japonica (Laminariales, Phaeophyceae)
摘要
Abstract
Phosphomannomutase (PMM) is an important enzyme involved in the synthesis of fucoidan and alginate. Two PMM genes of Saccharina japonica (Sjpmm1 and Sjpmm2) were cloned by rapid-amplification of cDNA ends (RACE). The open reading frame (ORF) length of Sjpmm1 is 759 bp, encoding 252 amino acids (SjPMM1) with a molecular weight (MW) of 28.51 kDa which belongs to the haloacid dehalogenase (HAD) superfamily. While the ORF length of Sjpmm2 is 1866 bp, encoding 621 amino acids (SjPMM2) with a MW of 66.49 kDa which belongs to the phosphohexomutase superfamily. The α-helix is the major secondary structure for both SjPMM1 and SjPMM2. The phylogenetic analysis showed that Sjpmm1 evolved from ancient eukaryotes, while Sjpmm2 originated from primary endosymbiosis. In addition, real-time PCR analysis indicated that temperature could increase the transcrip-tional level of Sjpmm, which may lead to the upregulation of fucoidan. Furthermore, a high concentration of the SjPMM1 fusion protein was expressed with the pMAL-c5X vector, contributing to the further function studies.关键词
海带/磷酸甘露糖变位酶/岩藻聚糖/褐藻胶/实时定量 PCR 分析Key words
Saccharina japonica/phosphomannomutase/fucoidan/alginate/real-time PCR analysis分类
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张朋艳,于雪,姚建亭,段德麟..海带磷酸甘露糖变位酶（PMM）基因的克隆与表达分析[J].海洋科学,2016,40(3):32-39,8.基金项目
国家科技支撑计划项目(2013BAB01B01) （2013BAB01B01）
国家海洋公益性行业科研专项(201405040)@@@@National Key Technology Research and Development Pro-gram (2013BAB01B01) （201405040）
Ocean Public Welfare Scientific Research Project (201405040) （201405040）
