Identification and cloning of C. albicans SC5314 genes encoding L-methionine biosynthetic pathway enzymes. - Open Research Data - Bridge of Knowledge

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Identification and cloning of C. albicans SC5314 genes encoding L-methionine biosynthetic pathway enzymes.

Description

Enzymes of fungal L-methionine biosynthetic pathway: homoserine O-acetyltransferase (Met2p), O-acetylhomoserine sulfhydrylase (Met15p) and cystathionine-γ-synthase (Str2p) could be exploited as molecular targets for antifungal chemotherapy. The goal of the study was to identify and clone genes encoding mentioned above enzymes. MET2, MET15 and STR2 genes were identified with the use of Candida Genome Database (http://www.candidagenome.org/). Amplifications of genes were carried out on C. albicans SC5314 genomic DNA with the use of polymerase chain reaction (PCR) and appropriate designed primers. PCR products that give satisfying molecular weight were selected and then cloned in a vector for its maintenance to produce the target protein. The constructed expression plasmids were designed in three different versions: encoding a native protein or recombinants with oligo-His tags added either at the C- or N-end. Introduction of fusion domains would facilitate future isolation of the gene products via metal-affinity chromatography. The additional oligo-His domains were introduced by PCR with the help of appropriate designed primers. A direct mutagenesis was carried out in a case of the plasmid containing MET2 gene because it contains a codon CTG that is translated differently in bacteria and yeast cells. In C. albicans it encodes L-Serine, whereas in E. coli cells L-Leucine.
This dataset contains the description of methodology, used primers sequences and three image files with the results of agarose gel electrophoresis of obtained PCR products, agarose gel electrophoresis of isolated plasmids DNA and restriction analysis.
The studies were financially supported from the project OPUS 20 financed by the National Science Centre (No. UMO-2020/39/B/NZ7/01519).

Dataset file

Idenification and cloning of C. albicans MET2, MET15, STR2 genes.zip
1.8 MB, S3 ETag 66b50b2c079cc24b385bc67f92658599-1, downloads: 9
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download file Idenification and cloning of C. albicans MET2, MET15, STR2 genes.zip

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License:
Creative Commons: by 4.0 open in new tab
CC BY
Attribution
Raw data:
Data contained in dataset was not processed.

Details

Year of publication:
2022
Verification date:
2022-01-18
Dataset language:
English
Fields of science:
  • Chemical sciences (Natural sciences)
DOI:
DOI ID 10.34808/zac2-tz70 open in new tab
Funding:
Verified by:
Gdańsk University of Technology

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