Cloning of the Rhizopus niveus pyr4 gene and its use for the transformation of Rhizopus delemar.
We have cloned a pyr4 gene encoding orotidine-5'-monophosphate decarboxylase of the filamentous fungus Rhizopus niveus. The pyr4 gene of R. nivens has an open reading frame composed of 265 amino-acid residues and has two putative introns. We have also isolated a pyr4 mutant of Rhizopus delemar from 5-fluoroorotic acid-resistant mutants and transformed it with the pyr4 gene of R. niveus as a selectable marker. Introduced DNA was integrated into the chromosome in a multiple tandem array. The mitotic stability of the introduced DNA was increased by a repeated sporulation process. The expression of the Escherichia coli beta-glucuronidase gene in R. delemar was successfully obtained under the control of the pgk2 gene promoter of R. niveus by co-transformation with the pyr4 gene.
Horiuchi
H
H
Department of Biotechnology, University of Tokyo, Japan.
Takaya
N
N
Yanai
K
K
Nakamura
M
M
Ohta
A
A
Takagi
M
M
D000595:Amino Acid Sequence; D001483:Base Sequence; D002262:Carboxy-Lyases; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000235:genetics*; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D005819:Genetic Markers; D008969:Molecular Sequence Data; D009154:Mutation; D012233:Rhizopus / Q000145:classification / Q000235:genetics*; D014170:Transformation, Genetic
D005656:Fungal Proteins; D005819:Genetic Markers; D002262:Carboxy-Lyases; C096137:pyr4 protein, Rhizopus