Title: Evaluation of a candidate control gene for Flavobacterium columnare expression analysis and expression profiles in four growth media Authors
Submitted to: Book of Abstracts Aquaculture America
Publication Type: Abstract Only
Publication Acceptance Date: September 23, 2010
Publication Date: March 2, 2011
Citation: Farmer, B.D., Straus, D.L., Fuller, S.A. 2011. Evaluation of a candidate control gene for Flavobacterium columnare expression analysis and expression profiles in four growth media [abstract]. Aquaculture America 2011: Aquaculture on Parade, February 28-March 3, 2011, New Orleans, Lousiana. p.147. Technical Abstract: Investigations into gene expression often yield important data that cannot be ascertained by other techniques. Studies of this nature hinge strongly on the reference or control gene. Primers and a fluorescent probe were designed to amplify the 16S ribosomal gene of Flavobacterium columnare. The 16S primer/probe set was compared to the existing QPCR primer/probe set of the chondrotinase gene; both genes were amplified under identical conditions with identical components. Results showed that both primer/probe sets had very similar efficiency (1.97 vs. 1.94) and cycle variation (CV). Genes were expressed in media as well as in infected fish tissues. Standard procedures for the RT PCR of the genes for basic relative expression ('' CT) has been tested using two isolates. Gene expression was profiled after isolates were cultured in various media. This data can be used to validate the 16S primer/probe set, stability of the control gene and optimal media.