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首页> 外文期刊>Protein Expression and Purification >Screening molluscan cDNA expression libraries with anti-shell matrix antibodies
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Screening molluscan cDNA expression libraries with anti-shell matrix antibodies

机译：用抗壳基质抗体筛选molluscan cDNA表达文库

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In a previous paper [Marin et al., Protein Expr. Purif. 23 (2001) 175], we showed that polyclonal antibodies raised against molluscan shell matrices could be useful tools for visualizing shell proteins after a preparative fractionation of the shell matrix. In this paper, we have used the same antibodies for screening a cDNA library constructed from mantle tissues of the nacro-prismatic bivalve Pinna nobilis. The immunoscreening led to the identification of a new protein, mucoperlin [Marin et al., J. Biol. Chem. 275 (2000) 20667], which was subsequently overexpressed. A polyclonal antibody was obtained from the recombinant mucoperlin. In a control assay, we unambiguously demonstrated that this antibody and one of the sera used for the initial screening hybridize with the same clones. We assess that screening cDNA libraries with antibodies elicited against unfractionated calcifying matrices is a good alternative to oligonucleotide screening techniques, particularly in the field of molluscan biomineralization where only few gene sequences are known. (C) 2003 Elsevier Science (USA). All rights reserved. [References: 22]
机译：在以前的论文中[Marin等，Protein Expr。 Purif。 23（2001）175]，我们显示了针对软体动物壳基质的多克隆抗体可能是有用的工具，用于在壳基质的制备分离后可视化壳蛋白。在本文中，我们使用了相同的抗体筛选了由棱角棱双壳双壳Pinna nobilis的地幔组织构建的cDNA文库。免疫筛选导致鉴定了新蛋白粘蛋白perlin [Marin等人，J.Biol。化学275（2000）20667]，随后被过分表达。从重组粘菌素获得多克隆抗体。在对照试验中，我们明确证明该抗体与用于初始筛选的血清之一与相同克隆杂交。我们评估了用针对未分级的钙化基质的抗体筛选cDNA文库是寡核苷酸筛选技术的良好选择，特别是在软体动物生物矿化领域中，只有很少的基因序列已知。 （C）2003 Elsevier Science（美国）。版权所有。 [参考：22]

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