The complete karyotype of Aspergillus niger: the use of introduced electrophoretic mobility variation of chromosomes for gene assignment studies.
A method is described for unambiguous assignment of cloned genes to Aspergillus niger chromosomes by CHEF gel electrophoresis and Southern analysis. All of the eight linkage groups (LGs), with the exception of LG VII, have previously been assigned to specific chromosomal bands in the electrophoretic karyotype of A. niger. Using a LG VII-specific probe (nicB gene of A. niger) we have shown that LG VII corresponds to a chromosome of about 4.1 Mb. Furthermore, genetic localization of three unassigned genes (glaA, aglA and pepA) in strains in which these genes had been replaced by a selectable marker gene led to a revised karyotype for the chromosomes corresponding to LGs VIII and VI. The revised electrophoretic karyotype reveals only 5 distinct bands. The presence of three pairs of equally sized chromosomes precluded assignment of genes to one specific chromosome in the wild-type strain. However, unambiguous chromosome assignment of cloned genes using CHEF-Southern analysis was demonstrated using a set of A. niger strains with introduced chromosomal size variation. The availability of these tester strains obviates the need to isolate or construct mutant strains for the purpose of chromosome assignment.
Verdoes
J C
JC
Department of Molecular Genetics and Gene Technology, TNO Nutrition and Food Research Institute, Rijswijk, The Netherlands.
Calil
M R
MR
Punt
P J
PJ
Debets
F
F
Swart
K
K
Stouthamer
A H
AH
van den Hondel
C A
CA
D001234:Aspergillus niger / Q000235:genetics*; D002874:Chromosome Mapping; D015825:Chromosomes, Fungal; D004587:Electrophoresis, Agar Gel; D008040:Genetic Linkage; D007621:Karyotyping