合浦珠母贝matrilin-1基因的克隆和表达分析

Molecular cloning and expression profiles of matrilin-1 in pearl oyster (Pinctada fucata)

  • 摘要: 以合浦珠母贝(Pinctada fucata)转录组中的matrilin-1基因片段为基础,开展了matrilin-1的cDNA全长(Pfmatrilin-1)克隆和定量表达分析。Pfmatrilin-1基因cDNA全长2 036 bp,开放阅读框(open reading frame,ORF)1 194 bp,编码397个氨基酸,包括1个信号肽序列和2个血管性血友病因子A(von Willebrand factor A,VWA)样结构域。系统进化分析显示无脊椎动物的matrilins与脊椎动物的matrilins进化关系较远。荧光定量PCR表达分析结果显示,Pfmatrilin-1 mRNA在合浦珠母贝各个组织中均有表达,其中在血液中表达量最高(P < 0.05);Pfmatrilin-1从担轮幼虫期至变态期均有表达,眼点期表达量最高(P < 0.05),而贝类幼虫发育到眼点期就会开始附着,眼点期幼虫原壳生长停滞,次生壳形成,因此推测其可能与次生壳的形成有关。这些结果表明Pfmatrilin-1在合浦珠母贝生物矿化中发挥重要作用。

     

    Abstract: Matrilin-1 gene was cloned from the Pinctada fucata based on its transcriptome sequences and named as Pfmatrilin-1. The whole length of Pfmatrilin-1 was 2 036 bp, including a 1 194 bp ORF encoding 397 amino acids, in which there were a signal peptide and two VWA (von Willebrand factor A) motifs. Phylogeny analysis proves that the matrilins in vertebrates and invertebrates were of distant relationship. The qRT-PCR tissue expression analysis indicates that Pfmatrilin-1 was expressed in all the tested tissues with significantly high expression level in blood cells (P < 0.05).The Pf-matrilin-1 was also continuously expressed from trochophore to metamorphosis stages, reaching the peak at pediveliger stage (P < 0.05).The prodissoconchⅡformed during pediveliger stage, so the Pfmatrilin-1 may be involved in regulating the formation of prodissoconchⅡ. Thus, it is proved that the Pfmatrilin-1 plays an important role of biomineralization in P.fucata.

     

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