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

Ann N Y Acad Sci. 2005 Apr:1040:301-4. doi: 10.1196/annals.1327.047.

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

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Animals
  • Cichlids / genetics
  • Cichlids / metabolism*
  • Gene Expression Regulation / physiology
  • Growth Hormone / analysis
  • Growth Hormone / biosynthesis
  • Growth Hormone / genetics*
  • Insulin-Like Growth Factor I / analysis
  • Insulin-Like Growth Factor I / biosynthesis
  • Insulin-Like Growth Factor I / genetics*
  • Insulin-Like Growth Factor II / analysis
  • Insulin-Like Growth Factor II / biosynthesis
  • Insulin-Like Growth Factor II / genetics*
  • Reverse Transcriptase Polymerase Chain Reaction / methods*

Substances

  • Insulin-Like Growth Factor I
  • Insulin-Like Growth Factor II
  • Growth Hormone