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Abstract: We have employed targeted gene cloning via PCR and differential screening of cDNA libraries to clone morphogenes in Agaricus bisporus. Using degenerate PCR primers an ca. 680 bp chitin synthase (CHS) gene fragment was amplified from A. bisporus. A button stage cDNA library was screened, with the CHS gene fragment, and the full length cDNA (chsl) was cloned. Sequence analysis revealed an ORF of 2727 bases encoding a putative peptide of 909 amino acids. Northern analysis revealed considerably enhanced chsl gene transcript in the fruit body. Differential screening of a cDNA library from the veil break stage mushrooms (stage 4), with the cDNA probe from button stage mushrooms (stage 2) yielded 32 up-regulated clones. Preliminary sequencing categorised the clones into 10 putative genes or gene families and Northern analysis confirmed their up-regulation. Further sequencing and analysis of the pattern of regulation of the genes are taking place.
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