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Fungal Genetics Reports Volume 37 Article 20 Linkage mapping of the gpdA gene of Aspergillus nidulans P. J. PUNT Medical Biological Laboratory TNO P. A. GREAVES Medical Biological Laboratory TNO CAM J. J. VAN DEN HONDEL Medical Biological Laboratory TNO Follow this and additional works at: http://newprairiepress.org/fgr Recommended Citation PUNT, P. J., P.A. GREAVES, and C.J. VAN DEN HONDEL (1990) "Linkage mapping of the gpdA gene of Aspergillus nidulans," Fungal Genetics Reports: Vol. 37, Article 20. https://doi.org/10.4148/1941-4765.1485 This Regular Paper is brought to you for free and open access by New Prairie Press. It has been accepted for inclusion in Fungal Genetics Reports by an authorized administrator of New Prairie Press. For more information, please contact [email protected]. Linkage mapping of the gpdA gene of Aspergillus nidulans Abstract In the last few years many genes of several Aspergillus species have been cloned and sequenced. For many of these genes mutant alleles and genetic linkage data are also available. However, for those genes for which no mutant alleles have been isolated, genetic mapping was not possible. Here we report linkage mapping of the glyceraldehyde-3- phosphate dehydrogenase gene (gpdA) of A. nidulans for which no mutant alleles have been isolated. The method used is applicable to all other cloned genes. Creative Commons License This work is licensed under a Creative Commons Attribution-Share Alike 4.0 License. This regular paper is available in Fungal Genetics Reports: http://newprairiepress.org/fgr/vol37/iss1/20 Linkage mapping of the gpdA gene of Aspergillus nidulans P.J. Punt, P.A. Greaves and C.A.M.J.J. van den Hondel - Medical Biological Laboratory TNO, Lange Kleiweg 139, P.O. Box 45, 2280 AA Rijswijk, The Netherlands. In the last few years many genes of several Aspergillus species have been cloned and sequenced. For many of these genes mutant alleles and genetic linkage data are also available. However, for those genes for which no mutant alleles have been isolated, genetic mapping was not possible. Here we report linkage mapping of the glyceraldehyde-3- phosphate dehydrogenase gene (gpdA) of A. nidulans for which no mutant alleles have been isolated. The method used is applicable to all other cloned genes. Transformation in Aspergillus frequently occurs by homologous recombination between host and vector sequences. Although the frequency of homologous recombination does not need to be identical for different sequences, a vector containing sequences of the gene to be mapped will often integrate at the chromosomal locus of this gene. In this way, A. nidulans ArgB[pAN541B]15 was obtained (vector pAN5-41B contains the lacZ gene fused to the promoter region of the gpdA gene of A. nidulans ; Van Gorcom et al. 1986 Gene 48:211-217). Southern blot analysis has shown that this strain contains a single copy of a (functional) lacZ gene at the gpdA locus. Parasexual analysis of this strain with master strain MSE (A. nidulans FGSC A288) was carried out. Segregation of the lacZ marker (integrated at the gpdA locus) was analysed (Table I). Table I. Linkage group assignment FGSC A288 gpdA:lacZ(a) I yA2 62/64 * ArgB[pAN5-41B]15 biA1 gpdA:lacZ 63/138 46% II III wA3 galA1 2/139 47/121 1% 39% - argB2 ** - IV V pyroA4 facA303 72/135 63/139 53% 45% VI sB3 ND ND methG2 66/138 48% - - VII VIII nicB8 riboB2 61/133 59/138 45% 43% - - a - the number and % of recombinants is given; two independent diploids were analysed. The markers located at both arms of chromosome IV (pyroA4 and methG2) gave 25/133 (19%) recombinants in this experiment. * - wA3 is epistatic to yA2 ** - only ArgB+ segregants were analysed From the results in Table I we conclude that the lacZ gene is significantly linked to wA3 at chromosome II: thus, gpdA is located on chromosome II. Using suitable linkage group II strains the location of gpdA on chromosome II can be mapped similarly, although the presence of duplicated sequences in ArgB[pAN5-41B]15 could disturb normal segregation in sexual crosses. Published by New Prairie Press, 2017