Cloning, expression and characterization of <em>Pinctada maxima</em> mantle gene 1

WANG Yu-mei; XIA Jian-hong; HUANG Gui-ju; YU Da-hui

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WANG Yu-mei, XIA Jian-hong, HUANG Gui-ju, YU Da-hui. Cloning, expression and characterization of Pinctada maxima mantle gene 1[J]. Haiyang Xuebao, 2010, 32(3): 121-128.

Citation:

WANG Yu-mei, XIA Jian-hong, HUANG Gui-ju, YU Da-hui. Cloning, expression and characterization of Pinctada maxima mantle gene 1[J]. Haiyang Xuebao, 2010, 32(3): 121-128.

WANG Yu-mei, XIA Jian-hong, HUANG Gui-ju, YU Da-hui. Cloning, expression and characterization of Pinctada maxima mantle gene 1[J]. Haiyang Xuebao, 2010, 32(3): 121-128.

Citation:

WANG Yu-mei, XIA Jian-hong, HUANG Gui-ju, YU Da-hui. Cloning, expression and characterization of Pinctada maxima mantle gene 1[J]. Haiyang Xuebao, 2010, 32(3): 121-128.

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Cloning, expression and characterization of Pinctada maxima mantle gene 1

1.
Key Laboratory of Ministry of Agriculture for Mariculture, Ecology and Quality Control, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, China;College of Fisheries and Life Science, Shanghai Ocean Univer
2.
Key Laboratory of Ministry of Agriculture for Mariculture, Ecology and Quality Control, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, China

Received Date: 2009-04-13
Rev Recd Date: 2010-03-10

Abstract

Abstract

EF-hand motif plays essential roles in the absorption and transportation of calcium ion in eukaryotic cells, and is possibly involved in the formation of shell and nacre in oysters. Degenerated primers were designed according to the conserved sequences of reported EF-hand proteins in oysters, and a novel gene PMMG1(Pinctada maxima mantle gene 1) was screened from the mantle cDNA library of P. maxima. The PMMG1 cDNA contained 618 nucleotides and encoded 140 amino acids (aa), including a putative signal peptide of 22 aa. PMMG1 protein shared an identity of 56％ with PFMG1 from P. fucata and had two putative EF-hand motifs. The cDNA fragment encoding mature protein of PMMG1 was cloned and integrated into prokaryotic expression vector pET32-a. A recombinant protein of expected size was induced by IPTG and then purified by Ni²⁺-NTA resin. Electrophoretic shift experiment revealed that the PMMG1 protein could bind both Ca²⁺ and Mg²⁺ion, and the tissue-specific expression level of PMMG1 was much higher in mantle than in other tissues, which may be involved in the formation of calcite while binding Ca²⁺ or the formation of aragonite while binding Mg²⁺. This work can benefit the further investigation into the roles of EF-hand proteins in the biomineralization of pearl oysters.
- Pinctada maxima,
- PMMG1,
- EF-hand domain,
- recombinant protein,
- nacre

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References(40)

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