The advantage of absolute quantification in comparative hormone research as indicated by a newly established real-time RT-PCR: GH, IGF-I, and IGF-II gene expression in the tilapia, Oreochromis niloticus.
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.
Eppler
Elisabeth
E
Division of Neuroendocrinology, Institute of Anatomy, University of Zürich, Switzerland.
Caelers
Antje
A
Berishvili
Giorgi
G
Reinecke
Manfred
M
D000818:Animals; D023681:Cichlids / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000502:physiology; D013006:Growth Hormone / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D007334:Insulin-Like Growth Factor I / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D007335:Insulin-Like Growth Factor II / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
D007334:Insulin-Like Growth Factor I; D007335:Insulin-Like Growth Factor II; D013006:Growth Hormone