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High-throughput identification of heavy metal binding proteins from the byssus of chinese green mussel (Perna viridis) by combination of transcriptome and proteome sequencing

机译:转录组和蛋白质组测序相结合高通量鉴定青贻贝(Perna viridis)的重金属结合蛋白

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摘要

The Byssus, which is derived from the foot gland of mussels, has been proved to bind heavy metals effectively, but few studies have focused on the molecular mechanisms behind the accumulation of heavy metals by the byssus. In this study, we integrated high-throughput transcriptome and proteome sequencing to construct a comprehensive protein database for the byssus of Chinese green mussel (Perna viridis), aiming at providing novel insights into the molecular mechanisms by which the byssus binds to heavy metals. Illumina transcriptome sequencing generated a total of 55,670,668 reads. After filtration, we obtained 53,047,718 clean reads and subjected them to de novo assembly using Trinity software. Finally, we annotated 73,264 unigenes and predicted a total of 34,298 protein coding sequences. Moreover, byssal samples were analyzed by proteome sequencing, with the translated protein database from the foot transcriptome as the reference for further prediction of byssal proteins. We eventually determined 187 protein sequences in the byssus, of which 181 proteins are reported for the first time. Interestingly, we observed that many of these byssal proteins are rich in histidine or cysteine residues, which may contribute to the byssal accumulation of heavy metals. Finally, we picked one representative protein, Pvfp-5-1, for recombinant protein synthesis and experimental verification of its efficient binding to cadmium (Cd2+) ions.
机译:来自贻贝脚腺的比索斯已被证明可以有效地结合重金属,但是很少有研究关注比索积累重金属的分子机制。在这项研究中,我们整合了高通量的转录组和蛋白质组测序,以构建一个综合的蛋白质数据库,用于中国青贻贝(Perna viridis)的囊泡,旨在为囊泡与重金属结合的分子机制提供新颖的见解。 Illumina转录组测序共产生了55,067,668个读段。过滤后,我们获得了53,047,718个干净的读数,并使用Trinity软件对其进行了重新组装。最后,我们注释了73,264个单基因,并预测了总共34,298个蛋白质编码序列。此外,通过蛋白质组测序分析了附睾样品,并将来自脚转录组的翻译蛋白质数据库作为进一步预测附睾蛋白质的参考。我们最终确定了囊中的187个蛋白序列,其中首次报道了181个蛋白。有趣的是,我们观察到许多这些byssal蛋白富含组氨酸或半胱氨酸残基,这可能有助于重金属的bysal积累。最后,我们选择了一种代表性蛋白Pvfp-5-1进行重组蛋白合成,并通过实验验证了其与镉(Cd 2 + )离子的有效结合。

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