Direct Amplification of an α-Amylase Gene from Barley Chromosome 6 by a Microdissection Method

概要

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Laser microdissection and subsequent direct DNA amplification methods are both effective for plant genome research. By using these techniques, we isolated barley chromosome 6 and amplified the target gene, a-amyl. Barley a-amyl gene, encoding the high-pI a-amylase, has been mapped onto the long arm of chromosome 6 by Southern hybridization and isozyme analysis using lines with specific chromosome constitution. The primers based on the a-amyl sequence were used for PCR with the isolated chromosomes. When the five microisolated chromosome 6 were used as the template, expected fragments with 142 bp were amplified. By contrast, no PCR product was detected when five chromosome 7 were used as a negative control. As a result, we directly confirmed that the a-amyl gene locates on barley chromosome 6, and concluded that five microisolated chromosomes are sufficient for direct amplification of the target gene.
上越教育大学の論文