13 research outputs found
Isolation, characterisation and expression patterns of a RAD51 ortholog from Pleurotus ostreatus
No full textAB: Using degenerated primers for conserved regions of RecA homologs we have isolated a gene from Pleurotus ostreatus that shows characteristic features of RAD51 homologs. The encoded amino acid sequence of P. ostreatus RAD51 (PoRAD51) shows greatest sequence similarities with RAD51 from Coprinus cinereus (89% identity). Furthermore the genomic organisation of PoRAD51 is almost identical to that of RAD51 from C. cinereus. Northern analysis shows that the expression of PoRAD51 is found in vegetative mycelium, and fruit body tissue, and that it is expressed at elevated levels in lamellae/basidia and following DNA damage. A sporulation deficient mutant strain of P. ostreatus (ATTC 58937) showed expression patterns of the RAD51 gene that are similar those of the normal sporulating strain
Isolation, characterization, and expression patterns of a DMC1 homolog from the basidiomycete Pleurotus ostreatus
No full textHere we describe the isolation of a Pleurotus ostreatus gene PoDMC1. The predicted amino acid sequence of the oyster mushroom gene is 62% identical to the yeast DMC1 and 60% identical to human DMC1. The highest degree of amino acid identity (88%), however, was shown with Coprinus CoLIM15, a DMC1 homolog recently found in Coprinus cinereus. The exact matching of sizes and positions of most introns in both basidiomycete genes underlines the close relationship between these DMC1 orthologs. The RecA homolog DMC1 from yeast and its orthologs from other species have been reported to be meiosis specific and essential for sporulation. Here we show that PoDMC1 is exclusively expressed in the lamellae/basidiospore fraction of fruit bodies and not in somatic cells of fruiting bodies or in vegetative mycelium. Furthermore, the gene is not expressed in the lamellae/basidiospore fraction of a nonsporulating mutant of P. ostreatus. Since one of the major problems in cultivating the oyster mushroom is the abundant sporulation that causes allergic reactions in man, PoDMC1 could be an important target gene in constructing sporeless Pleurotus strain
Isolation, characterization, and expression patterns of a DMC1 homolog from the basidiomycete Pleurotus ostreatus
No full textHere we describe the isolation of a Pleurotus ostreatus gene PoDMC1. The predicted amino acid sequence of the oyster mushroom gene is 62% identical to the yeast DMC1 and 60% identical to human DMC1. The highest degree of amino acid identity (88%), however, was shown with Coprinus CoLIM15, a DMC1 homolog recently found in Coprinus cinereus. The exact matching of sizes and positions of most introns in both basidiomycete genes underlines the close relationship between these DMC1 orthologs. The RecA homolog DMC1 from yeast and its orthologs from other species have been reported to be meiosis specific and essential for sporulation. Here we show that PoDMC1 is exclusively expressed in the lamellae/basidiospore fraction of fruit bodies and not in somatic cells of fruiting bodies or in vegetative mycelium. Furthermore, the gene is not expressed in the lamellae/basidiospore fraction of a nonsporulating mutant of P. ostreatus. Since one of the major problems in cultivating the oyster mushroom is the abundant sporulation that causes allergic reactions in man, PoDMC1 could be an important target gene in constructing sporeless Pleurotus strain
Transformation of the cultivated mushroom Agaricus bisporus (Lange) using T-DNA from Agrobacterium tumefaciens.
No full textAgrobacterium tumefaciens is known to transfer parts of its tumour-inducing plasmid, the T-DNA, to plants, yeasts and filamentous fungi. We have used this system to transform germinating basidiospores and vegetative mycelium of a commercial strain of the cultivated basidiomycete Agaricus bisporus. Analysis of transformants shows that the T-DNA integrates at random sites into the host genome and that the selection marker is stable during mitosis and meiosis. The Agrobacterium system allows the transformation of both homokaryons and heterokaryons of A. bisporus. Also, both karyotypes of an heterokaryon can be transformed simultaneously. Furthermore, this is the first report on the transformation of vegetative mycelium of a commercial strain of A. bisporus
Isolation, characterisation and expression patterns of a RAD51 ortholog from Pleurotus ostreatus
No full textAB: Using degenerated primers for conserved regions of RecA homologs we have isolated a gene from Pleurotus ostreatus that shows characteristic features of RAD51 homologs. The encoded amino acid sequence of P. ostreatus RAD51 (PoRAD51) shows greatest sequence similarities with RAD51 from Coprinus cinereus (89% identity). Furthermore the genomic organisation of PoRAD51 is almost identical to that of RAD51 from C. cinereus. Northern analysis shows that the expression of PoRAD51 is found in vegetative mycelium, and fruit body tissue, and that it is expressed at elevated levels in lamellae/basidia and following DNA damage. A sporulation deficient mutant strain of P. ostreatus (ATTC 58937) showed expression patterns of the RAD51 gene that are similar those of the normal sporulating strain
Abr1, a transposon-like element in the genome of the cultivated mushroom Agaricus bisporus (Lange) Imbach
No full textAbr1, a transposon-like element in the genome of the cultivated mushroom Agaricus bisporus (Lange) Imbach
No full textMicroRNA-29 is an essential regulator of brain maturation through regulation of CH methylation
No full textAcknowledgments We thank the members of the Deshmukh Lab for critical review of this manuscript. We also acknowledge Dr. Natallia Riddick, Viktoriya Nikolova, and Dr. Kara Agster at the UNC Mouse Behavioral Phenotyping Laboratory, for their technical assistance. We thank Mervi Eeva, Ying Li, and Bentley Midkiff at the UNC Translational Pathology Laboratory for expert technical assistance. We also appreciate the technical assistance provided by Janice Weaver and Carolyn Suitt at the UNC Animal Histopathology and the Center for Gastrointestinal Biology and Disease (CGIBD), respectively. The graphical abstract was created partly with BioRender.com. This work was supported by NIH, United States (GM118331 and AG055304 to M.D.). H.S. is a Howard Hughes Medical Institute Fellow of the Damon Runyon Cancer Research Foundation, United States (DRG-2194-14). J.M.S. and T.S.P. were supported by The Eunice Kennedy Shriver National Institute of Child Health and Human Development (U54HD079124) and NINDS (P30NS045892) of United States. The UNC Mouse Behavioral Phenotyping Laboratory is supported by a grant from the National Institute of Child Health and Human Development (NICHD), United States (U54-HD079124). The UNC Translational Pathology Laboratory is supported in part by grants from the NCI (5P30CA016086-42), NIH (U54-CA156733), NIEHS (5 P30 ES010126-17), UCRF, and NCBT (2015-IDG-1007) of United States. Author contributions V.S., A.N., and E.H. conducted most of the experiments with help as described here. H.S. and M.E.G. performed the whole-genome bisulfite sequencing and its analyses. J.M.S. and T.S.P. analyzed all RNA-seq data. C.L.K., M.K., P.S., and S.M.H. performed small RNA-seq and miRHub analysis. C.P., J.G., and E.A. helped with imaging and analysis of immunohistochemical stains. C.F. and M.B. helped manage the mouse colony. J.L. and Y.-W.H. generated the miR-29-floxed mice. S.M. conducted and analyzed many of the neurobehavioral assessments. V.S., M.E.G., and M.D. outlined the project. M.D. supervised the project. V.S., A.N., E.H., and M.D. produced the final version of the manuscript.Peer reviewe
