Relationship of vector insert size to homologous integration during transformation of Neurospora crassa with the cloned am (GDH) gene

1990 ◽  
Vol 221 (1) ◽  
pp. 37-43 ◽  
Author(s):  
David K. Asch ◽  
John A. Kinsey
Keyword(s):  
Neurospora Crassa ◽  
Insert Size ◽  
Homologous Integration ◽  
Relationship Of

Transformation of Neurospora crassa with the cloned am (glutamate dehydrogenase) gene

1984 ◽  
Vol 4 (1) ◽  
pp. 117-122
Author(s):  
J A Kinsey ◽  
J A Rambosek
Keyword(s):  
Neurospora Crassa ◽  
Glutamate Dehydrogenase ◽  
Plasmid Vector ◽  
Wild Type ◽  
Mendelian Segregation ◽  
Dehydrogenase Gene ◽  
Mutant Strains ◽  
Colonial Morphology ◽  
Relationship Of ◽  
Lambda Dna

We used DNA containing the am gene of Neurospora crassa, cloned in the lambda replacement vector lambdaL-47 (this clone is designated lambdaC-10), and plasmid vector subclones of this DNA to transform am deletion and point mutant strains. By means of subcloning, all sequences required for transformation to am prototrophy and expression of glutamate dehydrogenase have been shown to reside on a 2.5-kilobase BamHI fragment. We also characterized several am+ strains that were obtained after transformation with lambdaC-10. These strains showed Mendelian segregation of the am+ gene, although less than 50% of the transformed strains showed the normal linkage relationship of am with inl. In all cases tested, the strains had incorporated lambda DNA as well. The lambda DNA also showed a Mendelian segregation pattern. In one case, the incorporation of am DNA in a novel position was associated with a mutagenic event producing a strain with a very tight colonial morphology. In all cases in which the am+ gene had become the resident of a new chromosome, glutamate dehydrogenase was produced to only 10 to 20% of the wild-type levels.

scholarly journals Transformation of Neurospora crassa with the cloned am (glutamate dehydrogenase) gene.

1984 ◽  
Vol 4 (1) ◽  
pp. 117-122 ◽  
Author(s):  
J A Kinsey ◽  
J A Rambosek
Keyword(s):  
Neurospora Crassa ◽  
Glutamate Dehydrogenase ◽  
Plasmid Vector ◽  
Wild Type ◽  
Mendelian Segregation ◽  
Dehydrogenase Gene ◽  
Mutant Strains ◽  
Colonial Morphology ◽  
Relationship Of ◽  
Lambda Dna

We used DNA containing the am gene of Neurospora crassa, cloned in the lambda replacement vector lambdaL-47 (this clone is designated lambdaC-10), and plasmid vector subclones of this DNA to transform am deletion and point mutant strains. By means of subcloning, all sequences required for transformation to am prototrophy and expression of glutamate dehydrogenase have been shown to reside on a 2.5-kilobase BamHI fragment. We also characterized several am+ strains that were obtained after transformation with lambdaC-10. These strains showed Mendelian segregation of the am+ gene, although less than 50% of the transformed strains showed the normal linkage relationship of am with inl. In all cases tested, the strains had incorporated lambda DNA as well. The lambda DNA also showed a Mendelian segregation pattern. In one case, the incorporation of am DNA in a novel position was associated with a mutagenic event producing a strain with a very tight colonial morphology. In all cases in which the am+ gene had become the resident of a new chromosome, glutamate dehydrogenase was produced to only 10 to 20% of the wild-type levels.

The biosynthesis of the free sterols and sterol esters of Neurospora crassa

1979 ◽  
Vol 57 (2) ◽  
pp. 112-116 ◽  
Author(s):  
A. M. Pierce ◽  
H. D. Pierce Jr. ◽  
A. M. Unrau ◽  
A. C. Oehlschlager ◽  
R. E. Subden ◽  
...  
Keyword(s):  
Neurospora Crassa ◽  
Total Sterol ◽  
Mevalonic Acid ◽  
Ergosterol Biosynthesis ◽  
Sterol Esters ◽  
Free Sterols ◽  
Phase Culture ◽  
Esterified Sterols ◽  
Relationship Of ◽  
The Relationship

The composition of the free and esterified sterols in Neurospora crassa was examined as a function of incubation time in starvation medium containing [2-14C]mevalonic acid. The 14C incorporation was monitored in nuclear methylated and 4, 14-desmethyl sterol fractions. After 7 h incubation, sterol esterification had increased from an initial 5% in the log phase culture to 48% of the total sterol pool, with a concomitant decrease in free sterols. The relationship of the free and esterified sterol components in ergosterol biosynthesis is discussed.

scholarly journals Roles of the Mdm10, Tom7, Mdm12, and Mmm1 Proteins in the Assembly of Mitochondrial Outer Membrane Proteins in Neurospora crassa

2010 ◽  
Vol 21 (10) ◽  
pp. 1725-1736 ◽  
Author(s):  
Jeremy G. Wideman ◽  
Nancy E. Go ◽  
Astrid Klein ◽  
Erin Redmond ◽  
Sebastian W.K. Lackey ◽  
...  
Keyword(s):  
Neurospora Crassa ◽  
Outer Membrane ◽  
Double Mutant ◽  
Outer Membrane Proteins ◽  
Mitochondrial Outer Membrane ◽  
Mitochondrial Functions ◽  
Mitochondrial Distribution ◽  
Additive Loss ◽  
The Individual ◽  
Relationship Of

The Mdm10, Mdm12, and Mmm1 proteins have been implicated in several mitochondrial functions including mitochondrial distribution and morphology, assembly of β-barrel proteins such as Tom40 and porin, association of mitochondria and endoplasmic reticulum, and maintaining lipid composition of mitochondrial membranes. Here we show that loss of any of these three proteins in Neurospora crassa results in the formation of large mitochondrial tubules and reduces the assembly of porin and Tom40 into the outer membrane. We have also investigated the relationship of Mdm10 and Tom7 in the biogenesis of β-barrel proteins. Previous work showed that mitochondria lacking Tom7 assemble Tom40 more efficiently, and porin less efficiently, than wild-type mitochondria. Analysis of mdm10 and tom7 single and double mutants, has demonstrated that the effects of the two mutations are additive. Loss of Tom7 partially compensates for the decrease in Tom40 assembly resulting from loss of Mdm10, whereas porin assembly is more severely reduced in the double mutant than in either single mutant. The additive effects observed in the double mutant suggest that different steps in β-barrel assembly are affected in the individual mutants. Many aspects of Tom7 and Mdm10 function in N. crassa are different from those of their homologues in Saccharomyces cerevisiae.

scholarly journals An Improved Genome Assembly for Drosophila navojoa, the Basal Species in the mojavensis Cluster

2018 ◽  
Vol 110 (1) ◽  
pp. 118-123 ◽  
Author(s):  
Thyago Vanderlinde ◽  
Eduardo Guimarães Dupim ◽  
Nestor O Nazario-Yepiz ◽  
Antonio Bernardo Carvalho
Keyword(s):  
Genome Assembly ◽  
Chromosomal Rearrangements ◽  
Substantial Improvement ◽  
Insert Size ◽  
Genetic Studies ◽  
Cactophilic Drosophila ◽  
Host Shifts ◽  
Evolutionary Genetic ◽  
Relationship Of ◽  
Assembly Software

Abstract Three North American cactophilic Drosophila species, D. mojavensis, D. arizonae, and D. navojoa, are of considerable evolutionary interest owing to the shift from breeding in Opuntia cacti to columnar species. The 3 species form the “mojavensis cluster” of Drosophila. The genome of D. mojavensis was sequenced in 2007 and the genomes of D. navojoa and D. arizonae were sequenced together in 2016 using the same technology (Illumina) and assembly software (AllPaths-LG). Yet, unfortunately, the D. navojoa genome was considerably more fragmented and incomplete than its sister species, rendering it less useful for evolutionary genetic studies. The D. navojoa read dataset does not fully meet the strict insert size required by the assembler used (AllPaths-LG) and this incompatibility might explain its assembly problems. Accordingly, when we re-assembled the genome of D. navojoa with the SPAdes assembler, which does not have the strict AllPaths-LG requirements, we obtained a substantial improvement in all quality indicators such as N50 (from 84 kb to 389 kb) and BUSCO coverage (from 77% to 97%). Here we share a new, improved reference assembly for D. navojoa genome, along with a RNAseq transcriptome. Given the basal relationship of the Opuntia breeding D. navojoa to the columnar breeding D. arizonae and D. mojavensis, the improved assembly and annotation will allow researchers to address a range of questions associated with the genomics of host shifts, chromosomal rearrangements and speciation in this group.

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