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United States Department of Agriculture

Agricultural Research Service

Research Project: TERMITES: BIOLOGY, RISK ASSESSMENT AND SURVEILLANCE OF INVASIVE SPECIES Title: Screening of multiple potential control genes for use in caste and body region comparisons using RT-qPCR in Coptotermes formosanus

Authors
item Tarver, Matthew
item Mattison, Chris
item Florane, Christopher
item Hinchliffe, Doug
item Zhang, Dunhua
item Lax, Alan

Submitted to: Sociobiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: December 1, 2011
Publication Date: May 11, 2012
Citation: Tarver, M.R., Mattison, C.P., Florane, C.B., Hinchliffe, D.J., Zhang, D., Lax, A.R. 2012. Screening of multiple potential control genes for use in caste and body region comparisons using RT-qPCR in Coptotermes formosanus. Sociobiology. 59(1):81-96.

Interpretive Summary: Formosan subterranean termites, Coptotermes formosanus, are an important world wide pest. Molecular gene expression is an important tool for understanding the physiology of organisms. The recent advancement of molecular tools for Coptotermes formosanus is leading to the advancement of the understanding of termite physiology. One of the first steps in analyzing gene expression is the normalization to constant reference genes. Stable reference genes that have constant expression across multiple treatments are important for accurately comparing target genes expression. The objective of this investigation was to analyze and validate a set of potential reference genes including 18S rRNA; Glyceraldehyde 3-phosphate dehydrogenase (Gadphd); ribosomal protein L7 (RPL); ß-actin (BA1); a-tubulin (Atube); a-actin (Aactin); and elongation factor (Elong) as standards for analysis of transcriptional changes in the Formosan subterranean termite formosanus, across two phenotypic castes, body regions, and colonies. Our results demonstrate that 18S and RPL can serve as reliable expression standards when comparing these different castes and body regions. The results presented here help the future molecular studies of termite physiology.

Technical Abstract: Formosan subterranean termites, Coptotermes formosanus, are an important world wide pest. Molecular gene expression is an important tool for understanding the physiology of organisms. The recent advancement of molecular tools for Coptotermes formosanus is leading to advancement of the understanding of termite physiology. One of the first steps in analyzing gene expression is the normalization to constant reference genes. Stable reference genes that have constant expression across multiple treatments are important for accurately comparing target genes expression. The objective of this investigation was to analyze and validate a set of potential reference genes including 18S rRNA; Glyceraldehyde 3-phosphate dehydrogenase (Gadphd); ribosomal protein L7 (RPL); ß-actin (BA1); a-tubulin (Atube); a-actin (Aactin); and elongation factor (Elong) as standards for analysis of transcriptional changes in the termite Coptotermes formosanus, across two phenotypic castes, body regions, and colonies. We also compared the expression of hexamerin-1 and 2 using stable and unstable reference gene to demonstrate the importance of consistent control genes. Our results demonstrate that 18S and RPL can serve as reliable expression standards when comparing these different castes and body regions, and we show that C. formosanus Hex-1 and Hex-2 have expression patterns similar to that previously described in R. flavipes.

Last Modified: 7/24/2014
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