Development of an itraconazole resistance gene as a dominant selectable marker for transformation in Aspergillus oryzae and Aspergillus luchuensis

2021 ◽  
Vol 85 (3) ◽  
pp. 722-727
Author(s):  
Jikian Tokashiki ◽  
Hirohide Toyama ◽  
Osamu Mizutani
Keyword(s):  
Aspergillus Oryzae ◽  
Selectable Marker ◽  
Antifungal Drugs ◽  
Marker Genes ◽  
Resistance Marker ◽  
Synthesis Inhibitor ◽  
Fungal Cell ◽  
Steroid Synthesis ◽  
Dominant Selectable Marker ◽  
Aspergillus Luchuensis

ABSTRACT There are only a few combinations of antifungal drugs with known resistance marker genes in the Aspergillus species; therefore, the transformation of their wild-type strains is limited. In this study, to develop the novel dominant selectable marker for itraconazole, a fungal cell membrane synthesis inhibitor, we focused on Aspergillus luchuensis cyp51A (Alcyp51A), which encodes a 14-α-sterol demethylase related to the steroid synthesis pathway. We found that the G52R mutation in AlCyp51A and the replacement of the native promoter with a high-expression promoter contributed to itraconazole resistance in Aspergillus oryzae, designated as itraconazole resistant gene (itrA). The random integration in the A. luchuensis genome of the itrA marker cassette gene also allowed for transformation using itraconazole. Therefore, we succeed in developing a novel itraconazole resistance marker as a dominant selectable marker for transformation in A. oryzae and A. luchuensis.

Transformation of Aspergillus oryzae with a dominant selectable marker

1991 ◽  
Vol 19 (1) ◽  
pp. 117-122 ◽  
Author(s):  
S. Cheevadhanarak ◽  
G. Saunders ◽  
D.V. Renno ◽  
T.W. Flegel ◽  
G. Holt
Keyword(s):  
Aspergillus Oryzae ◽  
Selectable Marker ◽  
Dominant Selectable Marker

Cloning of the Ribosomal Protein L41 Gene of Phaffia rhodozyma and Its Use as a Drug Resistance Marker for Transformation

1998 ◽  
Vol 64 (5) ◽  
pp. 1947-1949 ◽  
Author(s):  
I.-G. Kim ◽  
S.-K. Nam ◽  
J.-H. Sohn ◽  
S.-K. Rhee ◽  
G.-H. An ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Ribosomal Protein ◽  
Copy Number ◽  
Selectable Marker ◽  
Phaffia Rhodozyma ◽  
Resistance Marker ◽  
Drug Resistance Marker ◽  
Targeting Signal ◽  
Dominant Selectable Marker ◽  
Dna Fragment

ABSTRACT The ribosomal protein L41 gene of Phaffia rhodozyma was cloned and used as a dominant selectable marker for cycloheximide resistance in the transformation of P. rhodozyma. Electrotransformation with a plasmid containing a ribosomal DNA fragment as a targeting signal typically yielded 800 to 1,200 transformants/μg of DNA with an integrated copy number of about seven per haploid genome.

Hygromycin B resistance as dominant selectable marker in yeast

1984 ◽  
Vol 8 (5) ◽  
pp. 353-358 ◽  
Author(s):  
Kevin R. Kaster ◽  
Stanley G. Burgett ◽  
Thomas D. Ingolia
Keyword(s):  
Selectable Marker ◽  
Hygromycin B ◽  
Hygromycin B Resistance ◽  
Dominant Selectable Marker

Insertional mutagenesis in Coprinus cinereus : use of a dominant selectable marker to generate tagged, sporulation-defective mutants

1999 ◽  
Vol 36 (6) ◽  
pp. 371-382 ◽  
Author(s):  
W. Jason Cummings ◽  
Martina Celerin ◽  
Jennifer Crodian ◽  
Linda K. Brunick ◽  
Miriam E. Zolan
Keyword(s):  
Insertional Mutagenesis ◽  
Selectable Marker ◽  
Coprinus Cinereus ◽  
Dominant Selectable Marker

The CRY1 gene in Chlamydomonas reinhardtii: structure and use as a dominant selectable marker for nuclear transformation

1994 ◽  
Vol 14 (6) ◽  
pp. 4011-4019
Author(s):  
J A Nelson ◽  
P B Savereide ◽  
P A Lefebvre
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Selectable Marker ◽  
Marker Gene ◽  
Small Subunit ◽  
Direct Selection ◽  
Nuclear Transformation ◽  
Ribulose Bisphosphate Carboxylase ◽  
Bisphosphate Carboxylase ◽  
Dominant Selectable Marker ◽  
Cry1 Gene

We have cloned and sequenced the CRY1 gene, encoding ribosomal protein S14 in Chlamydomonas reinhardtii, and found that it is highly similar to S14/rp59 proteins from other organisms, including mammals, Drosophila melanogaster, and Saccharomyces cerevisiae. We isolated a mutant strain resistant to the eukaryotic translational inhibitors cryptopleurine and emetine in which the resistance was due to a missense mutation (CRY1-1) in the CRY1 gene; resistance was dominant in heterozygous stable diploids. Cotransformation experiments using the CRY1-1 gene and the gene for nitrate reductase (NIT1) produced a low level of resistance to cryptopleurine and emetine. Resistance levels were increased when the CRY1-1 gene was placed under the control of a constitutive promoter from the ribulose bisphosphate carboxylase/oxygenase small subunit 2 (RBCS2) gene. We also found that the 5' untranslated region of the CRY1 gene was required for expression of the CRY1-1 transgene. Direct selection of emetine-resistant transformants was possible when transformed cells were first induced to differentiate into gametes by nitrogen starvation and then allowed to dedifferentiate back to vegetative cells before emetine selection was applied. With this transformation protocol, the RBCS2/CRY1-1 dominant selectable marker gene is a powerful tool for many molecular genetic applications in C. reinhardtii.

scholarly journals Bleomycin resistance: a new dominant selectable marker for plant cell transformation

1986 ◽  
Vol 7 (3) ◽  
pp. 171-176 ◽  
Author(s):  
Jacques Hille ◽  
Frank Verheggen ◽  
Peter Roelvink ◽  
Henk Franssen ◽  
Ab van Kammen ◽  
...  
Keyword(s):  
Plant Cell ◽  
Cell Transformation ◽  
Selectable Marker ◽  
Dominant Selectable Marker ◽  
Plant Cell Transformation
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