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Identification Of A Polyadenylated Transcript Correlated With Development In The Sexual Cycle Of Schizophyllum

C. Raper, W. Timberlake
Published 1985 · Biology

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Abstract A polyadenylated transcript of about 580 nucleotides has been determined to occur preferentially in the dikaryon of Schizophyllum commune. It was apparent in blots of RNA from the vegetative dikaryon (sexually differentiated cells which are normally capable of developing into fruiting bodies) and not apparent in blots of RNA from the two monokaryotic homokaryons (sexually nondifferentiated cells) that were mated to obtain the dikaryon. It was detected by probing gel blots of poly(A)+ RNA from the dikaryon and from each of the two component unmated monokaryons simultaneously with a subset of cDNAs specific to the dikaryon. The “dikaryon-specific” cDNAs were obtained by isolating poly(A)+ RNA from the dikaryon prior to fruiting, preparing 32P-labeled cDNA from this RNA, and selecting those cDNA sequences that did not hybridize to the poly(A)+ RNA of the two component monokaryons. A probe consisting of dikaryotic cDNA that did hybridize to the poly(A)+ RNA of the monokaryons served as the control. A relatively small degree of gene regulation at the transcriptional level appears to be involved in differentiation of the dikaryon from the component monokaryons. A series of successive hybridizations (“cascade hybridization”) of 32P-labeled cDNA from the dikaryon with poly(A)+ RNA from the monokaryons revealed only 2% of the starting material as noncomplementary to the RNA of the monokaryons. This is in marked contrast to the high degree of transcriptional regulation observed for developmental processes in other fungi such as Aspergillus nidulans.
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