title: Repetitive genomic sequences as a substrate for homologous integration in the Rhizopus oryzae genome.
abstract: The vast number of repetitive genomic elements was identified in the genome of Rhizopus oryzae. Such genomic repeats can be used as homologous regions for integration of plasmids. Here, we evaluated the use of two different repeats: the short (575 bp) rptZ, widely distributed (about 34 copies per genome) and the long (2053 bp) rptH, less prevalent (about 15 copies). The plasmid carrying rptZ integrated, but did so through a 2256-bp region of homology to the pyrG locus, a unique genomic sequence. Thus, the length of rptZ was below the minimal requirements for homologous strand exchange in this fungus. In contrast, rptH was used efficiently for homologous integration. The plasmid bearing this repeat integrated in multicopy fashion, with up to 25 copies arranged in tandem. The latter vector, pPyrG-H, could be a valuable tool for integration at homologous sequences, for such purposes as high-level expression of proteins.
authors.affiliation: Russian National Collection of Industrial Microorganisms (VKPM), State Research Institute of Genetics and Selection of Industrial Microorganisms (GosNIIgenetika), 1-st Dorozhny pr., 1, Moscow 117545, Russia. Electronic address: t_yuzbashev@genetika.ru.
authors.forename: Tigran V
authors.initials: TV
authors.lastname: Yuzbashev
authors.affiliation: Russian National Collection of Industrial Microorganisms (VKPM), State Research Institute of Genetics and Selection of Industrial Microorganisms (GosNIIgenetika), 1-st Dorozhny pr., 1, Moscow 117545, Russia.
authors.forename: Anna S
authors.initials: AS
authors.lastname: Larina
authors.affiliation: Russian National Collection of Industrial Microorganisms (VKPM), State Research Institute of Genetics and Selection of Industrial Microorganisms (GosNIIgenetika), 1-st Dorozhny pr., 1, Moscow 117545, Russia.
authors.forename: Tatiana V
authors.initials: TV
authors.lastname: Vybornaya
authors.affiliation: Russian National Collection of Industrial Microorganisms (VKPM), State Research Institute of Genetics and Selection of Industrial Microorganisms (GosNIIgenetika), 1-st Dorozhny pr., 1, Moscow 117545, Russia.
authors.forename: Evgeniya Y
authors.initials: EY
authors.lastname: Yuzbasheva
authors.affiliation: Russian National Collection of Industrial Microorganisms (VKPM), State Research Institute of Genetics and Selection of Industrial Microorganisms (GosNIIgenetika), 1-st Dorozhny pr., 1, Moscow 117545, Russia.
authors.forename: Ilia T
authors.initials: IT
authors.lastname: Gvilava
authors.affiliation: Russian National Collection of Industrial Microorganisms (VKPM), State Research Institute of Genetics and Selection of Industrial Microorganisms (GosNIIgenetika), 1-st Dorozhny pr., 1, Moscow 117545, Russia.
authors.forename: Sergey P
authors.initials: SP
authors.lastname: Sineoky
mesh_terms: D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D005822:Genetic Vectors; D005827:Genetics, Microbial / Q000379:methods; D016681:Genome, Fungal; D059765:Homologous Recombination; D008967:Molecular Biology / Q000379:methods; D008969:Molecular Sequence Data; D010957:Plasmids; D012091:Repetitive Sequences, Nucleic Acid; D012233:Rhizopus / Q000235:genetics*; D017422:Sequence Analysis, DNA; D014170:Transformation, Genetic
chemical_list: D004271:DNA, Fungal
keywords: Homologous recombination; Integrative transformation; Mucorales; Multicopy integration; Zygomycetes