[1]凌文康,黄慧炜,刘冰,等.贻贝足丝蛋白的汞、镉抗性及富集机制[J].深圳大学学报理工版,2019,36(No.5(473-600)):361-366.[doi:10.3724/SP.J.1249.2019.04361]
 LING Wenkang,HUANG Huiwei,LIU Bing,et al.Mercury and cadmium resistance and enrichment mechanism of mussel foot fibroin[J].Journal of Shenzhen University Science and Engineering,2019,36(No.5(473-600)):361-366.[doi:10.3724/SP.J.1249.2019.04361]
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贻贝足丝蛋白的汞、镉抗性及富集机制()
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《深圳大学学报理工版》[ISSN:1000-2618/CN:44-1401/N]

卷:
第36卷
期数:
2019年No.5(473-600)
页码:
361-366
栏目:
【生物工程】
出版日期:
2019-09-30

文章信息/Info

Title:
Mercury and cadmium resistance and enrichment mechanism of mussel foot fibroin
作者:
凌文康黄慧炜刘冰邓旭
深圳大学生命与海洋科学学院,广东深圳 518055
Author(s):
LING Wenkang HUANG Huiwei LIU Bing and DENG Xu
College of Life Sciences and Oceanography, Shenzhen University, Shenzhen 518055, Guangdong Province,P.R.China
关键词:
基因工程翡翠贻贝足丝蛋白基因克隆转录组学生物吸附重金属
Keywords:
genetic engineering Pernaviridis foot proteins transcriptomics biosorptionheavy metal
分类号:
Q78
DOI:
10.3724/SP.J.1249.2019.04361
文献标志码:
A
摘要:
为探究贻贝足丝蛋白黏附重金属的机理及分子机制,对翡翠贻贝足丝蛋白进行转录组测序分析,使用Trinity软件从头组装,获得73 571个UniGenes.翡翠贻贝足部的UniGenes注释到基因本体(gene ontology,GO)数据库,共获得9 466个注释结果,通过基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)数据库注释后,发现有955个UniGenes被注释到关于黏着斑蛋白的通路中,推测足丝蛋白吸附重金属的功能与其密切相关.由于Pvfp6半胱氨酸、组氨酸、酪氨酸含量高,该蛋白可能对重金属有结合作用.将pvfp6基因克隆到pET-30a表达载体,转到大肠杆菌BL21(DE3)表达,结果表明,含有pET-30a-pvfp6质粒的宿主具有一定的抗汞和抗镉能力,能分别在含Hg2+质量浓度为5 mg/L,或者含Cd2+质量浓度为160 mg/L的培养基中生长;重组菌对Hg2+和Cd2+的平衡富集量均高于对照组pET-30a,分别为109.89 mg/g和19.27 mg/g.研究表明,足丝蛋白Pvfp6赋予了重组菌具有一定的抗汞和抗镉的能力,且富集能力增强该研究可为海洋环境中重金属污染的生物治理奠定实验基础.
Abstract:
To study the molecular mechanism of mussel foot proteinadhering toheavy metal, 73571 UniGenes were derived by analyzing the transcriptome of Pernaviridis foot proteins and using Trinity assembler to construct denovo transcriptome assembly. After annotating to Gene Ontology, 9 466 UniGenes of Pernaviridis foot proteins were obtained. Further, 955 UniGenes were found to annotate to the focal adhesion pathway via KEGG. It is speculated that the function of foot proteins binding to heavy metal was closely related to this pathway. Concerning the high content of cysteine, histidine and tyrosine in Pvfp6, Pvfp6 may play an important role in heavy metal binding. Therefore, pvfp6 coding gene was cloned to pET-30a expression vector and transferred to E.coli BL21 (DE3). It turns out that the recombinant E.coli containingpET-30a-pvfp6 plasmid can grow in the presence of 5 mg/L Hg2+ or 160 mg/L Cd2+, showing stronger mercury and cadmium resistance than the wild type. Furthermore, the equilibrium accumulation capacities of Hg2+ and Cd2+ by the recombinant bacterial cells are 109.89mg/g and 19.27mg/g, respectively, higher than host cells.Summarily,the foot fibroin protein Pvfp6 improves not onlymercury and cadmium resistance of the recombinant bacteria, but also its bioaccumulation capacity. This study lays an experimental foundation for bioremediation of heavy metal pollution in marine environment.

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更新日期/Last Update: 2019-07-04