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United States Department of Agriculture

Agricultural Research Service

Title: Development of Ssh Libraries and Cdna Microarrays: Systems for Gene Expression Profiling and Functional Genomics of Greenbug

Author
item Huang, Yinghua

Submitted to: Society for Experimental Biology Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: June 15, 2005
Publication Date: July 11, 2005
Citation: Huang, Y. 2005. Development of SSH libraries and cDNA microarrays: systems for gene expression profiling and functional genomics of greenbug [abstract]. In: Comparative Biochemistry and Physiology. Annual Meeting of the Society for Experimental Biology, July 11-15, 2005, Barcelona, Spain. 141A(3):Suppl. p. S232.

Technical Abstract: The study of aphid-plant interactions can be greatly enhanced by the development of genomic tools for both an insect and its host plant. In this study, two subtractive cDNA libraries were constructed for greenbug using a PCR-based suppression subtractive hybridization (SSH) method. The two greenbug cDNA libraries were developed from two populations of mRNA derived from greenbugs feeding on a preferred host (referred as library 1) and a non-preferred host (library 2), respectively. Each cDNA population was enriched using the second cDNA population as the driver in cDNA subtraction. In addition, a normal unsubtracted cDNA library was also constructed to facilitate cloning of full-length cDNAs. Subsequently, high-throughput screening of the SSH cDNA libraries were preformed using DNA microarray analysis, in which a total of 2,304 selected cDNA clones represent 1536 unique transcripts from the library 1 and 768 from the library 2. In this way, special groups of differentially expressed genes were identified relevant to the feeding tests. The results generated from this study demonstrate that SSH technique and cDNA microarray are valuable tools for functional genomics of aphids. This powerful SSH technique has been able to compare two mRNA populations and obtained cDNAs of special genes that are either over-expressed or exclusively expressed in one population compared with another. This set of greenbug genes is an important resource for both identifying genes and gene regulation networks with key roles in compatible and incompatible plant-greenbug interactions and for understanding the genetic mechanisms underlying virulence of aphids and plant defense response.

Last Modified: 12/17/2014
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