A Microarray Approach To Identify The Differential Expression Patterns Of Barley Testa Specific Genes

Proanthocyanidins are prominent colorless compounds in the bark of trees, leaves, fruits, flowers and seed coats of many plant species. Proanthocyanindins are dimeric and polymeric condensation products of the flavonoid catechin, epicatechin or leucodelphinidin. Their toxicity deters insects and mammals from feeding on plants. Removals of proanthocyanidins from the barley seed coat has produced varieties that make the use of chemical stabilizers to achieve brilliant beer without chill-haze formation superfluous. In spite of numerous efforts the purification of leucocyanidin reductase converting (+)-2,3-trans-3,4-cis-leucocyanidin to (+)catechin and of an enzyme or an enzyme condensing leucocyanidin with catechin has not been achieved. In order to obtain the two enzymes, a molecular genetic approach is proposed.Recently, cDNA and microarrays have been developed and used to quantitate differential gene expression by hybridizing a complex mRNA-derived probe onto an array of PCR products representing specific cDNAs. Microarrays allow thousands of genes to be monitored simultaneously for expression level and compared between different materials.In this paper, we report: 1. Construction of wild type barley Morex testa+pericarp cDNA library, keep the cDNA fragments in phagemid. 2. Plating and titering some phagemid into E. coli strain SOLR, pick 64 plates of 96 well plate single colonies. 3. Print PCR amplified cDNA fragments on SuperAmine substrate. 4. Many differential expression cDNA fragments are found between wild type and mutant.