Bialaphos resistance as a dominant selectable marker in Neurospora crassa

1989 ◽  
Vol 16 (5-6) ◽  
pp. 369-372 ◽  
Author(s):  
Javier Avalos ◽  
Robert F. Geever ◽  
Mary E. Case
Keyword(s):  
Neurospora Crassa ◽  
Selectable Marker ◽  
Dominant Selectable Marker

scholarly journals Cloning and characterization of the gene for beta-tubulin from a benomyl-resistant mutant of Neurospora crassa and its use as a dominant selectable marker.

1986 ◽  
Vol 6 (7) ◽  
pp. 2452-2461 ◽  
Author(s):  
M J Orbach ◽  
E B Porro ◽  
C Yanofsky
Keyword(s):  
Neurospora Crassa ◽  
Selectable Marker ◽  
Resistant Mutant ◽  
Intron Position ◽  
Coding Region ◽  
Tubulin Gene ◽  
Beta Tubulin ◽  
Benomyl Resistance ◽  
Dominant Selectable Marker ◽  
Beta Tubulin Gene

We cloned the beta-tubulin gene of Neurospora crassa from a benomyl-resistant strain and determined its nucleotide sequence. The gene encodes a 447-residue protein which shows strong homology to other beta-tubulins. The coding region is interrupted by six introns, five of which are within the region coding for the first 54 amino acids of the protein. Intron position comparisons between the N. crassa gene and other fungal beta-tubulin genes reveal considerable positional conservation. The mutation responsible for benomyl resistance was determined; it caused a phenylalanine-to-tyrosine change at position 167. Codon usage in the beta-tubulin gene is biased, as has been observed for other abundantly expressed N. crassa genes such as am and the H3 and H4 histone genes. This bias results in pyrimidines in the third positions of 96% of the codons in codon families in which there is a choice between purines and pyrimidines in this position. Bias is also evident by the absence of 19 of the 61 sense codons. We demonstrated that benomyl resistance is due to the cloned beta-tubulin gene of strain Bml511(r)a and that this gene can be used as a dominant selectable marker in N. crassa transformation.

Cloning and characterization of the gene for beta-tubulin from a benomyl-resistant mutant of Neurospora crassa and its use as a dominant selectable marker

1986 ◽  
Vol 6 (7) ◽  
pp. 2452-2461
Author(s):  
M J Orbach ◽  
E B Porro ◽  
C Yanofsky
Keyword(s):  
Neurospora Crassa ◽  
Selectable Marker ◽  
Resistant Mutant ◽  
Intron Position ◽  
Coding Region ◽  
Tubulin Gene ◽  
Beta Tubulin ◽  
Benomyl Resistance ◽  
Dominant Selectable Marker ◽  
Beta Tubulin Gene

We cloned the beta-tubulin gene of Neurospora crassa from a benomyl-resistant strain and determined its nucleotide sequence. The gene encodes a 447-residue protein which shows strong homology to other beta-tubulins. The coding region is interrupted by six introns, five of which are within the region coding for the first 54 amino acids of the protein. Intron position comparisons between the N. crassa gene and other fungal beta-tubulin genes reveal considerable positional conservation. The mutation responsible for benomyl resistance was determined; it caused a phenylalanine-to-tyrosine change at position 167. Codon usage in the beta-tubulin gene is biased, as has been observed for other abundantly expressed N. crassa genes such as am and the H3 and H4 histone genes. This bias results in pyrimidines in the third positions of 96% of the codons in codon families in which there is a choice between purines and pyrimidines in this position. Bias is also evident by the absence of 19 of the 61 sense codons. We demonstrated that benomyl resistance is due to the cloned beta-tubulin gene of strain Bml511(r)a and that this gene can be used as a dominant selectable marker in N. crassa transformation.

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 Inactivation of the Neurospora crassa gene encoding the mitochondrial protein import receptor MOM19 by the technique of "sheltered RIP".

Genetics ◽  
1994 ◽  
Vol 136 (1) ◽  
pp. 107-118 ◽  
Author(s):  
T A Harkness ◽  
R L Metzenberg ◽  
H Schneider ◽  
R Lill ◽  
W Neupert ◽  
...  
Keyword(s):  
Neurospora Crassa ◽  
Target Gene ◽  
Protein Import ◽  
Selectable Marker ◽  
Mitochondrial Protein ◽  
Mutant Gene ◽  
Mitochondrial Protein Import ◽  
Gene Encoding ◽  
Precursor Proteins ◽  
Import Receptor

Abstract We have used a technique referred to as "sheltered RIP" (repeat induced point mutation) to create mutants of the mom-19 gene of Neurospora crassa, which encodes an import receptor for nuclear encoded mitochondrial precursor proteins. Sheltered RIP permits the isolation of a mutant gene in one nucleus, even if that gene is essential for the survival of the organism, by sheltering the nucleus carrying the mutant gene in a heterokaryon with an unaffected nucleus. Furthermore, the nucleus harboring the RIPed gene contains a selectable marker so that it is possible to shift nuclear ratios in the heterokaryons to a state in which the nucleus containing the RIPed gene predominates in cultures grown under selective conditions. This results in a condition where the target gene product should be present at very suboptimal levels and allows the study of the mutant phenotype. One allele of mom-19 generated by this method contains 44 transitions resulting in 18 amino acid substitutions. When the heterokaryon containing this allele was grown under conditions favoring the RIPed nucleus, no MOM19 protein was detectable in the mitochondria of the strain. Homokaryotic strains containing the RIPed allele exhibit a complex and extremely slow growth phenotype suggesting that the product of the mom-19 gene is important in N. crassa.

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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