Transformation of coccolithophorid <italic style="font-style: italic">Emiliania huxleyi</italic> harboring a marine virus (Coccolithoviruses) serine palmitoyltransferase (SPT) gene by electroporation

Weicong CAI; Xueting WANG; Jinjing SU; Jian LI; Jun ZENG; Guiling LI; Jingwen LIU

doi:10.1007/s00343-020-9325-0

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

Biology | Updated：2023-06-06

- Transformation of coccolithophorid Emiliania huxleyi harboring a marine virus (Coccolithoviruses) serine palmitoyltransferase (SPT) gene by electroporation
- Journal of Oceanology and Limnology Vol. 39, Issue 2, Pages: 693-704(2021)
- Affiliations：
  
  1.College of Food and Bioengineering, Jimei University, Xiamen 361021, China
  2.Fujian Provincial Key Laboratory of Food Microbiology and Enzyme Engineering, Xiamen 361021, China
- Author bio：
  
  Guiling LI, E-mail: guiling_li@hotmail.com
  Jingwen LIU, E-mail: ljwsbch@163.com
- Funds：
- DOI：10.1007/s00343-020-9325-0
  CLC：
- Received：17 December 2019，
  
  Accepted：17 March 2020，
  
  Online First：26 April 2020，
  
  Published：2021-03
- Accepted：
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Weicong CAI, Xueting WANG, Jinjing SU, et al. Transformation of coccolithophorid Emiliania huxleyi harboring a marine virus (Coccolithoviruses) serine palmitoyltransferase (SPT) gene by electroporation[J]. Journal of Oceanology and Limnology, 2021, 39(2): 693-704.
DOI：

Weicong CAI, Xueting WANG, Jinjing SU, et al. Transformation of coccolithophorid Emiliania huxleyi harboring a marine virus (Coccolithoviruses) serine palmitoyltransferase (SPT) gene by electroporation[J]. Journal of Oceanology and Limnology, 2021, 39(2): 693-704. DOI： 10.1007/s00343-020-9325-0.

摘要

Abstract

Emiliania huxleyi

is the most prominent modern coccolithophore

a group of marine unicellular eukaryotes that play a critical role in ocean biogeochemistry. Coccolithoviruses are large double stranded DNA viruses

which is responsible for the demise of large oceanic blooms formed by

E. huxleyi. E. huxleyi

virus (EhVs) acquired a series of enzyme-coding genes predicted to be involved in the sphingolipid biosynthesis by horizontal gene transfer between virus-host. Currently

there is limited experimental validation identifying the functions of these genes in EhV. Genetic transformation of eukaryotic cells is a powerful tool to get an insight into gene functions of the studied organisms. Serine palmitoyltransferase (SPT) catalyzes the first committed step in de novo sphingolipid biosynthetic pathway. Here

a novel vector system for the transformation of

E. huxleyi

was designed. It contained fragments of promoter and terminator sequences of

E. huxleyi

endogenic fucoxanthin chlorophyll

-binding protein gene "

fcp

" and harbored EhV-99B1

spt

gene. The resultant recombinant transformation vectors pEhux-I-

spt

and pEhux-II were co-transferred into

E. huxleyi

BOF92 by electroporation. Transformants were obtained upon glufosinate-ammonium selection

and confirmed by Southern hybridization

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 levels. The expression of the viral

spt

gene led to differences in lipid compositions analyzed using thin-layer chromatography (TLC). The results present the genetic transformation system for

E. huxleyi

providing additional genetic resource with potential for exploring basic biological questions such as the virus-host interactions.

关键词

Keywords

references

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Related Institution

College of Ocean Food and Biological Engineering, Jimei University

Key Laboratory of Marine Genetics and Breeding of Ministry of Education, Ocean University of China

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⁰

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

DOI：10.1007/s00343-020-9325-0

摘要

Abstract

关键词

Keywords

references