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Transformation of coccolithophorid Emiliania huxleyi harboring a marine virus(Coccolithoviruses) serine palmitoyltransferase(SPT) gene by electroporation

Transformation of coccolithophorid Emiliania huxleyi harbori...

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中国生物化学与分子生物学会第十二届全国会员代表大会暨2018年全国学术会议

作者： Cai Weicong Wang Xueting Guo Qiangqiang Li Guiling Zeng Jun Liu Jingwen College of Food and Bioengineering JiMei University

Emiliania huxleyi is the most prominent modern coccolithophore,a group of photosynthetic marine unicellular eukaryotes that play a critical role in ocean *** are large double stranded DNA viruses,which is responsible for the demise of large oceanic blooms formed *** virus acquired a series of enzyme-coding genes predicted to be involved in the sphingolipid biosynthesis by gene transfers between ***,there is limited experimental validation indentifying the functions of these *** palmitoyltransferase(spt) catalyzes the first committed step in de novo sphingolipid biosynthetic *** transformation of eukaryotic cells is a powerful tool to get an insight into gene functions of the studied ***,a novel vector system for the transformation of *** was *** contained larger fragments of promoter and terminator sequences of *** endogenic fucoxanthin chlorophyll a/c-binding protein gene "fcp" and harbored *** virus(EhV99 B1) spt *** resultant recombinant transformation vectors pEhux-I-spt and pEhux-II were co-transferred into *** BOF92 by *** were obtained upon glufosinate-ammonium selection,and confirmed by genome PCR,qRT-PCR and Western blot screening of spt gene,which indicated that spt gene was integrated into the nuclear genome and was expressed at the mRNA and protein *** expression of the viral spt gene led to the accumulation of lipid droplets in the transformed cells stained with Nile Red.

关键词： Emiliania huxleyi Coccllithovirus genetic transformation serine palmitoyltransferase

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Functional Inferences of Environmental Coccolithovirus Biodiversity

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Virologica Sinica 2013年第5期28卷 291-302页

作者： Jozef I Nissimov Mark Jones Johnathan A Napier Colin B Munn Susan A Kimmance Michael J Allen Plymouth Marine Laboratory Prospect Place University of Nottingham School of BiosciencesSutton Bonington Campus Department of Biological Chemistry Rothamsted Research School of Marine Science & Engineering Plymouth University

The cosmopolitan calcifying alga Emiliania huxleyi is one of the most abundant bloom forming coccolithophore species in the oceans and plays an important role in global biogeochemical cycling. Coccolithoviruses are a major cause of coccolithophore bloom termination and have been studied in laboratory, mesocosm and open ocean studies. However, little is known about the dynamic interactions between the host and its viruses, and less is known about the natural diversity and role of functionally important genes within natural coccolithovirus communities. Here, we investigate the temporal and spatial distribution of coccolithoviruses by the use of molecular fingerprinting techniques PCR, DGGE and genomie sequencing. The natural biodiversity of the virus genes encoding the major capsid protein （MCP） and serine palmitoyltransferase （SPT） were analysed in samples obtained from the Atlantic Meridional Transect （AMT）, the North Sea and the L4 site in the Westem Channel Observatory. We discovered nine new coccolithovirus genotypes across the AMT and L4 site, with the majority of MCP sequences observed at the deep chlorophyll maximum layer of the sampled sites on the transect. We also found four new SPT gene variations in the North Sea and at L4. Their translated fragments and the full protein sequence of SPT from laboratory strains EhV-86 and EhV-99B 1 were modelled and revealed that the theoretical fold differs among strains. Variation identified in the structural distance between the two domains of the SPT protein may have an impact on the catalytic capabilities of its active site. In summary, the combined use of ＇standard＇ markers （i.e. MCP）, in combination with metabolically relevant markers （i.e. SPT） are useful in the study of the phylogeny and functional biodiversity of coccolithoviruses, and can provide an interesting intracellular insight into the evolution of these viruses and their ability to infect and replicate within their algal hosts.

关键词： Coccolithovirus Major capsid protein serine palmitoyltransferase Functional biodiversity

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