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한국환경생물학회 / v.22, no.2, 2004년, pp.329-335 
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무당개구리 비텔로제닌 유전자의 발현의 RT- PCR 검출법
( RT- PCR Analysis of Vitellogenin Gene Expression in Bombina orientalis ) |
| 계명찬;이명식;강희정;정경아;안혜선; 한양대학교 자연과학대학 생명과학과;한양대학교 자연과학대학 생명과학;한양대학교 자연과학대학 생명과학;한양대학교 자연과학대학 생명과학;한양대학교 자연과학대학 생명과학과;
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| 초 록 |
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| 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. |
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| 키워드 |
| vitellogenin;RT- PCR;liver;biomarker;endocrine disrupter;Bombina orientalis; |
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환경생물 / v.22, no.2, 2004년, pp.329-335
한국환경생물학회
ISSN : 1226-9999
UCI : G100:I100-KOI(KISTI1.1003/JNL.JAKO200411922362343)
언어 : 한국어 |
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| 논문 제공 : KISTI 한국과학기술정보연구원 |
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