4.4 Article

Establishment of a new cell line from the snout tissue of golden pompano Trachinotus ovatus, and its application in virus susceptibility

Journal

JOURNAL OF FISH BIOLOGY
Volume 88, Issue 6, Pages 2251-2262

Publisher

WILEY
DOI: 10.1111/jfb.12986

Keywords

cell line; snout; transgenic manipulation; virus infection

Funding

  1. National High Technology Development Program of China [2014AA093507]
  2. National Basic Research Program of China [2012CB114402]
  3. Science and Technology and industrial development Marine fishery Project of Guangdong Province, China [A201501C01]
  4. Key Laboratory of Tropical Marine Bio-resources and Ecology, Chinese Academy of Sciences [LMB141005]

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A new marine-fish cell line, designated GPS, was established from the snout tissue of golden pompano Trachinotus ovatus. GPS cells multiplied well in Leibovitz's L-15 containing 10% foetal bovine serum (FBS) at 28 degrees C and the cells have been subcultured for >60 passages. Polymerase chain reaction (PCR) amplification of 16S ribosomal (r) RNA confirmed the origin of this cell line from T. ovatus. Chromosome analysis showed that GPS cells exhibited chromosomal aneuploidy with a modal chromosome number of 54. Bright green fluorescence signal was observed in enhanced green fluorescent protein (EGFP)-N3 transfected cells, indicating that GPS cells could be used to investigate gene functions in vitro. The GPS cells were highly susceptible to Singapore grouper iridovirus (SGIV), which was demonstrated by the presence of severe cytopathic effect (CPE) and increased viral titres. Real-time quantitative PCR and Western blot analysis showed that the viral gene transcription and protein synthesis occurred during SGIV infection in GPS cells. Thus, this study described the characteristic of a new cell line from the snout tissue of T. ovatus that could be used as a tool for propagation of iridovirus and genetic manipulation to investigate host-pathogen interactions. (C) 2016 The Fisheries Society of the British Isles

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