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海洋浮游植物中组蛋白基因的分子克隆、转录组分析及特征鉴定

Molecular Cloning, Transcriptome Profiling, and Characterization of Histone Genes in the Dinoflagellate .

作者信息

Riaz Sadaf, Sui Zhenghong

机构信息

Key Laboratory of Marine Genetics and Breeding (Ocean University of China), Ministry of Education, Qingdao, 266003, P.R. China.

Department of Microbiology, University of Veterinary and Animal Sciences, Lahore, Pakistan.

出版信息

J Microbiol Biotechnol. 2018 Jul 28;28(7):1185-1198. doi: 10.4014/jmb.1802.01075.

Abstract

The nucleosomal organization of chromatin using histone proteins is a fundamental and ubiquitous feature of eukaryotic nuclei, with the major exception of dinoflagellates. Although a number of recent genomic and transcriptomic analyses have detected numerous histone genes in dinoflagellates, little is known about their expression. Here in, we aimed to investigate the expression pattern of histone genes under nutritional stress, and an attempt was made to detect histone expression at the protein level in . The presence of histones at the mRNA level was confirmed in this study by the amplification, cloning, and sequencing of 10 different genes. Relative expression profiling of these genes under different growth conditions was determined with real-time PCR and revealed considerable levels of histone transcription in nutritionally stressed cells. We were unable to detect the expression of histones at the protein level even after immunodetection and analysis using mass spectrometry, although a histone-like protein was detected as a major nuclear component. expresses multiple variants of histone, and protein sequences revealed both conservation and divergence with respect to other eukaryotes. We concluded that maintained an active transcription of histone genes within the cell, and enhanced expression of histone genes in nutritional stress strongly suggest that histones have functional significance in dinoflagellates, although expression at the protein level was below our current detection limits, which suggests a limited role of histones in DNA packaging. Finally, the plausible regulation of histone expression at the gene and protein levels in is discussed.

摘要

利用组蛋白形成的染色质核小体结构是真核细胞核的一个基本且普遍存在的特征,除了甲藻纲生物是主要的例外情况。尽管最近的一些基因组和转录组分析在甲藻纲生物中检测到了大量组蛋白基因,但对其表达情况却知之甚少。在此,我们旨在研究营养胁迫下组蛋白基因的表达模式,并尝试在[具体生物名称未给出]中检测蛋白质水平的组蛋白表达。在本研究中,通过对10个不同基因的扩增、克隆和测序,证实了组蛋白在mRNA水平的存在。利用实时PCR测定了这些基因在不同生长条件下的相对表达谱,结果显示在营养胁迫的细胞中组蛋白转录水平相当高。尽管检测到一种类组蛋白作为主要的核成分,但即使经过免疫检测和质谱分析,我们仍无法在蛋白质水平检测到组蛋白的表达。[具体生物名称未给出]表达多种组蛋白变体,其蛋白质序列显示出与其他真核生物既有保守性又有差异。我们得出结论,[具体生物名称未给出]在细胞内维持组蛋白基因的活跃转录,营养胁迫下组蛋白基因表达增强强烈表明组蛋白在甲藻纲生物中具有功能意义,尽管蛋白质水平的表达低于我们目前的检测限,这表明组蛋白在DNA包装中的作用有限。最后,讨论了[具体生物名称未给出]中组蛋白在基因和蛋白质水平上可能的调控机制。

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