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KMID : 1025520040460030315
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Journal of Animal Science and Technology
2004 Volume.46 No. 3 p.315 ~ p.324
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Detection of Tissue-specific Expression of Porcine Cytochrome P450 Aromatase Genes by Use of Denaturing High Performance Liquid Chromatography(DHPLC) Technique
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Chae S. H.
Ghimeray Amal Kumar
Hong J. M.
Lee E. J.
Chang J. S.
Choi In-Ho
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Abstract
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Cytochrome P450 aromatase is the enzyme responsible for biosynthesis of female sex hormone(estrogen) and 19-nortestosterone(nandrolone), a unique steroid hormone endogenously synthesized in the pig. By use of RT-PCR coupled with DHPLC technique (WAVE analysis), expression pattern of isoforms of porcine cytochrome P450 aromatase gene was investigated. Relatively higher expression of aromatase mRNA was observed in testis than in ovary and this result accounted for the previous findings of higher blood estrogen level in male compared with female in this species. The result from the DHPLC demonstrated that PCR amplified DNA fragments of ovary and testis tissues, using unique PCR primers for all three types of aromatase genes, were different from those of type Ⅱ and Ⅲ genes. Further nucleotide sequence analyses of the plasmid clones containing the PCR products revealed that nucleotide sequences of all clones were identical to type I aromatase gene(ovary type). Thus, the result from the present study indicates that the ovary and testis express the same type of aromatase gene. Therefore, the efficacy of DHPLC techniques used for this study helped us to analyze tissue-specific expression of isoform of genes containing the nucleotide sequences with high homology.
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KEYWORD
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Aromatase, DHPLC, Anabolic steroid, Testosterone, 19-Nortestosterone, Estrogen
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