Abstract
We have developed a real-time RT-PCR that absolutely quantifies the gene expression of hormones using the standard curve method. The method avoids cloning procedures by using primer extension to create templates containing a T7 promoter gene sequence. It is rapid since neither separate reverse transcriptions nor postamplification steps are necessary, and its low detection level (2 pg/mug total RNA) allows precise absolute quantification. Using the method, we have quantified the gene expression of GH, IGF-I, and IGF-II in the tilapia.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
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Review
MeSH terms
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Animals
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Cichlids / genetics
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Cichlids / metabolism*
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Gene Expression Regulation / physiology
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Growth Hormone / analysis
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Growth Hormone / biosynthesis
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Growth Hormone / genetics*
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Insulin-Like Growth Factor I / analysis
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Insulin-Like Growth Factor I / biosynthesis
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Insulin-Like Growth Factor I / genetics*
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Insulin-Like Growth Factor II / analysis
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Insulin-Like Growth Factor II / biosynthesis
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Insulin-Like Growth Factor II / genetics*
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Reverse Transcriptase Polymerase Chain Reaction / methods*
Substances
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Insulin-Like Growth Factor I
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Insulin-Like Growth Factor II
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Growth Hormone