Korean Journal of Environmental Biology (환경생물)

Korean Society of Environmental Biology (한국환경생물학회)
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RT- PCR Analysis of Vitellogenin Gene Expression in Bombina orientalis

무당개구리 비텔로제닌 유전자의 발현의 RT- PCR 검출법

  • 계명찬 (한양대학교 자연과학대학 생명과학과) ;
  • 이명식 (한양대학교 자연과학대학 생명과학) ;
  • 강희정 (한양대학교 자연과학대학 생명과학) ;
  • 정경아 (한양대학교 자연과학대학 생명과학) ;
  • 안혜선 (한양대학교 자연과학대학 생명과학과)
  • Published : 2004.06.01
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Abstract

To develop a biomarker for the monitoring of the contamination of estrogenic endocrine disrupters in the aquatic environment, reverse transcription -polymerase chain reaction (RT-PCR) analysis of vitellogenin (Vg) mRNA expression was optimized in Bombina orientalis, a Korean red bellied toad species. Based on partial cDNA sequences of both Vg and beta actin genes of B. orientalis, specific primers for RT-PCR of Vg and beta actin mRNAs were developed. Semiquantitative RT-PCR of the Vg mRNA in liver was optimized using a beta actin mRNA as an internal control in both sexes. In female RT-PCR using $1\;\mu{g}$ of the liver cDNA resulted in a linear increment in the PCR product of Vg from 18 to 34 cycles of amplification. In male, on the contrary, the RT- PCR product was first detected at 30 cycles of amplification and a linear increment was observed from 30 to 40 cycles of amplification, suggesting that male B. orientalis expresses minute amount of Vg mRNA which is a $2^{-12}$ equivalent of female. In conclusion, the optimized protocol for semiquantitative RT-PCR analysis of Vg mRNA level in B. orientalis male liver will be useful for the environmental monitoring the xenoestrogen contamination in the freshwater environment in Korea.

Keywords

References

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