A novel vector element providing multicopy vector integration in Arxula adeninivorans.
An Arxula adeninivorans vector element has been identified that provides multicopy integration in an atrp1 host strain. The element consists of the ATRP1 selection marker fused to a newly generated truncated ALEU2 promoter of 53 bp. In the described example eight copies of an amyA expression vector encoding heterologous alpha-amylase from Bacillus amyloliquefaciens are integrated in the genome of the recombinant strain instead of a single copy observed when using the ATRP1 element with the complete promoter. The high copy number results in strains of superior productivity for a secreted recombinant alpha-amylase. The vector design enables the integration of a small vector fragment that consists of yeast DNA only providing high transformation frequencies and a high mitotic stability.
Steinborn
Gerhard
G
Leibniz Institut für Pflanzengenetik und Kulturpflanzenforschung, Gatersleben, Germany.
Gellissen
Gerd
G
Kunze
Gotthard
G
D019747:Aldose-Ketose Isomerases / Q000235:genetics; D001409:Bacillus cereus / Q000201:enzymology / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics; D018628:Gene Dosage; D005816:Genetic Complementation Test; D005822:Genetic Vectors / Q000235:genetics*; D008967:Molecular Biology / Q000379:methods*; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D004718:Saccharomycetales / Q000235:genetics*; D000516:alpha-Amylases / Q000096:biosynthesis / Q000235:genetics
D005656:Fungal Proteins; D000516:alpha-Amylases; D019747:Aldose-Ketose Isomerases