scholarly journals Characterization of an autonomously replicating sequence in Candida guilliermondii

2013 ◽  
Vol 168 (9) ◽  
pp. 580-588 ◽  
Author(s):  
Emilien Foureau ◽  
Vincent Courdavault ◽  
Sandra M. Navarro Gallón ◽  
Sébastien Besseau ◽  
Andrew J. Simkin ◽  
...  
Keyword(s):  
Candida Guilliermondii ◽  
Autonomously Replicating Sequence

Isolation and characterization of an autonomously replicating sequence from Ustilago maydis

1988 ◽  
Vol 8 (9) ◽  
pp. 3703-3709
Author(s):  
T Tsukuda ◽  
S Carleton ◽  
S Fotheringham ◽  
W K Holloman
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Consensus Sequence ◽  
Ustilago Maydis ◽  
Small Fragment ◽  
Direct Repeats ◽  
Base Pairs ◽  
Autonomously Replicating Sequence ◽  
Isolation And Characterization ◽  
Extrachromosomal Elements

DNA fragments that function as autonomously replicating sequences (ARSs) have been isolated from Ustilago maydis. When inserted into an integrative transforming vector, the fragments increased the frequency of U. maydis transformation several-thousandfold. ARS-containing plasmids were transmitted in U. maydis as extrachromosomal elements through replication. They were maintained at a level of about 25 copies per cell but were mitotically unstable. One ARS characterized in detail, which we called UARS1, was localized to a 1.7-kilobase fragment. UARS1 contained a cluster of active sequences. This element could be reduced further into three separate subfragments, each of which retained ARS activity. The smallest one was 383 base pairs (bp) long. Although not active itself in yeast, this small fragment contained seven 8-bp direct repeats, two contiguous 30-bp direct repeats, and five 11-bp units in both orientations with sequences similar but not identical to the consensus sequence found to be crucial for ARS activity in Saccharomyces cerevisiae.

scholarly journals Candida guilliermondii Complex Is Characterized by High Antifungal Resistance but Low Mortality in 22 Cases of Candidemia

2017 ◽  
Vol 61 (7) ◽  
Author(s):  
Laura Judith Marcos-Zambrano ◽  
Mireia Puig-Asensio ◽  
Felipe Pérez-García ◽  
Pilar Escribano ◽  
Carlos Sánchez-Carrillo ◽  
...  
Keyword(s):  
Metabolic Activity ◽  
Biofilm Formation ◽  
Galleria Mellonella ◽  
Sensu Stricto ◽  
Candida Guilliermondii ◽  
Molecular Techniques ◽  
Content Type ◽  
Content Model ◽  
Crystal Violet Assay

ABSTRACT The objectives of our study were to describe the characteristics of patients with Candida guilliermondii candidemia and to perform an in-depth microbiological characterization of isolates and compare them with those of patients with C. albicans candidemia. We described the risk factors and outcomes of 22 patients with candidemia caused by the C. guilliermondii complex. Incident isolates were identified using molecular techniques, and susceptibility to fluconazole, anidulafungin, and micafungin was studied. Biofilm formation was measured using the crystal violet assay (biomass production) and the XTT reduction assay (metabolic activity), and virulence was studied using the Galleria mellonella model. Biofilm formation was compared with that observed for C. albicans. The main conditions predisposing to infection were malignancy (68%), immunosuppressive therapy (59%), and neutropenia (18%). Clinical presentation of candidemia was less severe in patients infected by the C. guilliermondii complex than in patients infected by C. albicans, and 30-day mortality was lower in C. guilliermondii patients (13.6% versus 33.9%, respectively; P = 0.049). Isolates were identified as C. guilliermondii sensu stricto (n = 17) and Candida fermentati (n = 5). The isolates produced biofilms with low metabolic activity and moderate biomass. The G. mellonella model showed that C. guilliermondii was less virulent than C. albicans (mean of 6 days versus 1 day of survival, respectively; P < 0.001). Patients with candidemia caused by the C. guilliermondii complex had severe and debilitating underlying conditions. Overall, the isolates showed diminished susceptibility to fluconazole and echinocandins, although poor biofilm formation and the low virulence were associated with a favorable outcome.

scholarly journals Isolation and characterization of sequences from mouse chromosomal DNA with ARS function in yeasts.

1983 ◽  
Vol 3 (11) ◽  
pp. 1898-1908 ◽  
Author(s):  
G E Roth ◽  
H M Blanton ◽  
L J Hager ◽  
V A Zakian
Keyword(s):  
Cell Lines ◽  
Mouse Tissue ◽  
Chromosomal Dna ◽  
Autonomously Replicating Sequence ◽  
Culture Cells ◽  
Isolation And Characterization ◽  
Multiple Copies ◽  
Mouse Cells ◽  
Simplex Virus

Fragments of chromosomal DNA from a variety of eucaryotes can act as ARSs (autonomously replicating sequence) in yeasts. ARSs enable plasmids to be maintained in extrachromosomal form, presumably because they function as initiation sites for DNA replication. We isolated eight different sequences from mouse chromosomal DNA which function as ARSs in Saccharomyces cerevisiae (bakers' yeast). Although the replication efficiency of the different mouse ARSs in yeasts appears to vary widely, about one-half of them functions as well as the yeast chromosomal sequence ARS1. Moreover, five of the ARSs also promote self replication of plasmids in Schizosaccharomyces pombe (fission yeast). Each of the ARSs was cloned into plasmids suitable for transformation of mouse tissue culture cells. Plasmids were introduced into thymidine kinase (TK)-deficient mouse L cells by the calcium phosphate precipitation technique in the absence of carrier DNA. In some experiments, the ARS plasmid contained the herpes simplex virus type 1 TK gene; in other experiments (cotransformations), the TK gene was carried on a separate plasmid used in the same transformation. In contrast to their behavior in yeasts, none of the ARS plasmids displayed a significant increase in transformation frequency in mouse cells compared with control plasmids. Moreover, only 1 of over 100 cell lines contained the original plasmid in extrachromosomal form. The majority of cell lines produced by transformation with an ARS TK plasmid contained multiple copies of plasmid integrated into chromosomal DNA. In most cases, results with plasmids used in cotransformations were similar to those for plasmids carrying TK. However, cell lines produced by cotransformations with plasmids containing any one of three of the ARSs (m24, m25, or m26) often contained extrachromosomal DNAs.

scholarly journals Isolation and characterization of an autonomously replicating sequence from Ustilago maydis.

1988 ◽  
Vol 8 (9) ◽  
pp. 3703-3709 ◽  
Author(s):  
T Tsukuda ◽  
S Carleton ◽  
S Fotheringham ◽  
W K Holloman
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Consensus Sequence ◽  
Ustilago Maydis ◽  
Small Fragment ◽  
Direct Repeats ◽  
Base Pairs ◽  
Autonomously Replicating Sequence ◽  
Isolation And Characterization ◽  
Extrachromosomal Elements

DNA fragments that function as autonomously replicating sequences (ARSs) have been isolated from Ustilago maydis. When inserted into an integrative transforming vector, the fragments increased the frequency of U. maydis transformation several-thousandfold. ARS-containing plasmids were transmitted in U. maydis as extrachromosomal elements through replication. They were maintained at a level of about 25 copies per cell but were mitotically unstable. One ARS characterized in detail, which we called UARS1, was localized to a 1.7-kilobase fragment. UARS1 contained a cluster of active sequences. This element could be reduced further into three separate subfragments, each of which retained ARS activity. The smallest one was 383 base pairs (bp) long. Although not active itself in yeast, this small fragment contained seven 8-bp direct repeats, two contiguous 30-bp direct repeats, and five 11-bp units in both orientations with sequences similar but not identical to the consensus sequence found to be crucial for ARS activity in Saccharomyces cerevisiae.

scholarly journals Characterization of xylose reductase extracted by CTAB-reversed micelles from Candida guilliermondii homogenate

2006 ◽  
Vol 42 (2) ◽  
pp. 251-257
Author(s):  
Ely Vieira Cortez ◽  
Adalberto Pessoa-Jr ◽  
Maria das Graças de Almeida Felipe ◽  
Inês Conceição Roberto ◽  
Michele Vitolo
Keyword(s):  
Xylose Reductase ◽  
Candida Guilliermondii ◽  
Reversed Micelles

Location and characterization of autonomously replicating sequences from chromosome VI of Saccharomyces cerevisiae

1993 ◽  
Vol 13 (8) ◽  
pp. 5043-5056
Author(s):  
K Shirahige ◽  
T Iwasaki ◽  
M B Rashid ◽  
N Ogasawara ◽  
H Yoshikawa
Keyword(s):  
Essential Element ◽  
The Other ◽  
Flanking Sequence ◽  
Functional Elements ◽  
Autonomously Replicating Sequence ◽  
The Core ◽  
Initiation Of Replication ◽  
Flanking Region ◽  
Delta G

We have reported the isolation of linking clones of HindIII and EcoRI fragments, altogether spanning a 230-kb continuous stretch of chromosome VI. The presence or absence of autonomously replicating sequence (ARS) activities in all of these fragments has been determined by using ARS searching vectors containing CEN4. Nine ARS fragments were identified, and their positions were mapped on the chromosome. Structures essential for and/or stimulative to ARS activity were determined for the ARS fragments by deletions and mutations. The organization of functional elements composed of core and stimulative sequences was found to be variable. Single core sequences were identified in eight of nine ARSs. The remaining ARS (ARS603) essential element is composed of two core-like sequences. The lengths of 3'- and 5'-flanking stimulative sequences required for the full activity of ARSs varied from ARS to ARS. Five ARSs required more than 100 bp of the 3'-flanking sequence as stimulative sequences, while not more than 79 bp of the 3' sequence was required by the other three ARSs. In addition, five ARSs had stimulative sequences varying from 127 to 312 bp in the 5'-flanking region of the core sequence. In general, these stimulative activities were correlated with low local delta Gs of unwinding, suggesting that the low local delta G of an ARS is an important element for determining the efficiency of initiation of replication of ARS plasmids.

scholarly journals Isolation and characterization of the centromere from chromosome V (CEN5) of Saccharomyces cerevisiae.

1984 ◽  
Vol 4 (1) ◽  
pp. 86-91 ◽  
Author(s):  
G T Maine ◽  
R T Surosky ◽  
B K Tye
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Sequence ◽  
Base Pair ◽  
Mapping Technique ◽  
Chromosome Loss ◽  
Tetrad Analysis ◽  
Autonomously Replicating Sequence ◽  
Centromeric Dna ◽  
Isolation And Characterization

We have cloned a functional centromeric DNA sequence from Saccharomyces cerevisiae. Using the 2 mu chromosome-loss mapping technique and meiotic tetrad analysis, we have identified this DNA sequence as the centromere of chromosome V (CEN5). The CEN5 sequence has been localized on an 1,100-base-pair BamHI-BglII restriction fragment. Plasmids containing CEN5 and an autonomously replicating sequence are mitotically stable in S. cerevisiae and segregate in a Mendelian fashion during meiosis.

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