Earticle

초록

영어

Loss of L-gulonolactone oxidase (GULO), which catalyzes the last step of the ascorbic acid (AA) biosynthesis pathway, results in a complete lack of AA in several Osteichthyes fish species, including zebrafish. In this study, sGULO, the active GULO gene from cloudy catshark (Scyliorhinus torazame) was cloned into zebrafish using the Gateway cloning method. The resulting Tg(b-actin:sGULO:mCherry) fish were analyzed for the effects of a reestablished AA pathway. Fluorescent microscopy and PCR were used to analyze the integration of the construct into the zebrafish genome. Catalytic activity of sGULO, AA production, growth-related characteristics, and gene expression were investigated to evaluate the effects of AA production in Tg fish. The mCherry fluorescent protein indicated the proper integration and expression of the sGULO construct in zebrafish. The sGULO gene was ubiquitously expressed in all the studied tissues and the enzyme activity indicated an increased AA production in Tg fish. The growth of Tg fish was also increased, and antioxidant system analysis suggests that reactive oxygen species production was reduced in Tg fish compared with wild type. Expression of the AA transporter slc23a1 was significantly downregulated in Tg homozygous fish. These results collectively indicate the effects of reestablished AA synthesis in zebrafish.

목차

Abstract
INTRODUCTION
MATERIALS AND METHODS
Zebrafish Husbandry
Bioinformatics Characterization and Isolation of Cloudy Catshark sGULO
Assembling the Expression Construct
Construction of Transgenic Zebrafish
Establishment of Transgenic Zebrafish
GULO Assay and AA Quantification
Feed Preparation
Growth Assay
Gene Expression Analysis
Statistical Analysis
RESULTS
Bioinformatics Analysis
Tg(b-actin:sGULO:mCherry) F0, F1, and F2 Generations
sGULO Enzyme Activity and Endogenous AA Production
Physiological Difference
Gene Expression Analysis
DISCUSSION
CONCLUSION
REFERENCES

키워드

저자

• K. A. S. N. Shanaka [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju, South Korea; Marine Science Institute, Jeju National University, Jeju, South Korea ]
• Sumi Jung [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju, South Korea; Marine Science Institute, Jeju National University, Jeju, South Korea ]
• N. D. Janson [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju, South Korea; Marine Science Institute, Jeju National University, Jeju, South Korea ]
• J. R. P. Jayasingha [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju, South Korea; Marine Science Institute, Jeju National University, Jeju, South Korea ]
• K. P. Madushani [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju, South Korea; Marine Science Institute, Jeju National University, Jeju, South Korea ]
• Myoung-Jin Kim [ Marine Science Institute, Jeju National University, Jeju, South Korea ] Corresponding Author
• Jehee Lee [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju, South Korea; Marine Science Institute, Jeju National University, Jeju, South Korea ] Corresponding Author

참고문헌

발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  해양과학연구소 연구논문집 [Bulletin of the Marine Science Institute]

• 간기

  폐간

• pISSN

  1225-5734

• 수록기간

  1977~2021

• 십진분류

  KDC 454 DDC 551