Construction of recombinant Kluyveromyces marxianus UFV-3 to express dengue virus type 1 nonstructural protein 1 (NS1)
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| 024 | 7 | 0 | |a 10.1007/s00253-014-5963-5 |2 doi |
| 035 | |a (NATIONALLICENCE)springer-10.1007/s00253-014-5963-5 | ||
| 245 | 0 | 0 | |a Construction of recombinant Kluyveromyces marxianus UFV-3 to express dengue virus type 1 nonstructural protein 1 (NS1) |h [Elektronische Daten] |c [Caio Bragança, Lívia Colombo, Alvaro Roberti, Mariana Alvim, Silvia Cardoso, Kledna Reis, Sérgio de Paula, Wendel da Silveira, Flavia Passos] |
| 520 | 3 | |a The yeast Kluyveromyces marxianus is a convenient host for industrial synthesis of biomolecules. However, despite its potential, there are few studies reporting the expression of heterologous proteins using this yeast. Here, we report expression of a dengue virus protein in K. marxianus for the first time. The dengue virus type 1 nonstructural protein 1 (NS1) was integrated into the K. marxianus UFV-3 genome at the LAC4 locus using an adapted integrative vector designed for high-level expression of recombinant protein in Kluyveromyces lactis. The NS1 gene sequence was codon-optimized to increase the level of protein expression in yeast. The synthetic gene was cloned in frame with K. lactis α-mating factor signal peptide, and the recombinant plasmid obtained was used to transform K. marxianus UFV-3 by electroporation. The transformed cells, selected in yeast extract peptone dextrose containing 200μgmL−1 Geneticin, were mitotically stable. Analysis of recombinant strains by RT-PCR and protein detection using blot analysis confirmed both transcription and expression of extracellular NS1 polypeptide. After induction with galactose, the NS1 protein was analyzed by sodium dodecyl sulfate-PAGE and immunogenic detection. Protein production was investigated under two conditions: with galactose and biotin pulses at 24-h intervals during 96h of induction and without galactose and biotin supplementation. Protease activity was not detected in post-growth medium. Our results indicate that recombinant K. marxianus is a good host for the production of dengue virus NS1 protein, which has potential for diagnostic applications. | |
| 540 | |a Springer-Verlag Berlin Heidelberg, 2014 | ||
| 690 | 7 | |a Kluyveromyces marxianus |2 nationallicence | |
| 690 | 7 | |a Heterologous expression |2 nationallicence | |
| 690 | 7 | |a Dengue virus |2 nationallicence | |
| 690 | 7 | |a NS1 protein |2 nationallicence | |
| 700 | 1 | |a Bragança |D Caio |u Laboratório de Fisiologia de Micro-organismos, Departamento de Microbiologia, BIOAGRO, Universidade Federal de Viçosa, Viçosa, MG, Brazil |4 aut | |
| 700 | 1 | |a Colombo |D Lívia |u Laboratório de Fisiologia de Micro-organismos, Departamento de Microbiologia, BIOAGRO, Universidade Federal de Viçosa, Viçosa, MG, Brazil |4 aut | |
| 700 | 1 | |a Roberti |D Alvaro |u Laboratório de Fisiologia de Micro-organismos, Departamento de Microbiologia, BIOAGRO, Universidade Federal de Viçosa, Viçosa, MG, Brazil |4 aut | |
| 700 | 1 | |a Alvim |D Mariana |u Laboratório de Fisiologia de Micro-organismos, Departamento de Microbiologia, BIOAGRO, Universidade Federal de Viçosa, Viçosa, MG, Brazil |4 aut | |
| 700 | 1 | |a Cardoso |D Silvia |u Laboratório de Imunovirologia Molecular, Departamento de Biologia Geral, Universidade Federal de Viçosa, Viçosa, MG, Brazil |4 aut | |
| 700 | 1 | |a Reis |D Kledna |u Microbiologia Veterinária Especial - MICROVET, Viçosa, MG, Brazil |4 aut | |
| 700 | 1 | |a de Paula |D Sérgio |u Laboratório de Imunovirologia Molecular, Departamento de Biologia Geral, Universidade Federal de Viçosa, Viçosa, MG, Brazil |4 aut | |
| 700 | 1 | |a da Silveira |D Wendel |u Laboratório de Fisiologia de Micro-organismos, Departamento de Microbiologia, BIOAGRO, Universidade Federal de Viçosa, Viçosa, MG, Brazil |4 aut | |
| 700 | 1 | |a Passos |D Flavia |u Laboratório de Fisiologia de Micro-organismos, Departamento de Microbiologia, BIOAGRO, Universidade Federal de Viçosa, Viçosa, MG, Brazil |4 aut | |
| 773 | 0 | |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/3(2015-02-01), 1191-1203 |x 0175-7598 |q 99:3<1191 |1 2015 |2 99 |o 253 | |
| 856 | 4 | 0 | |u https://doi.org/10.1007/s00253-014-5963-5 |q text/html |z Onlinezugriff via DOI |
| 898 | |a BK010053 |b XK010053 |c XK010000 | ||
| 900 | 7 | |a Metadata rights reserved |b Springer special CC-BY-NC licence |2 nationallicence | |
| 908 | |D 1 |a research-article |2 jats | ||
| 949 | |B NATIONALLICENCE |F NATIONALLICENCE |b NL-springer | ||
| 950 | |B NATIONALLICENCE |P 856 |E 40 |u https://doi.org/10.1007/s00253-014-5963-5 |q text/html |z Onlinezugriff via DOI | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Bragança |D Caio |u Laboratório de Fisiologia de Micro-organismos, Departamento de Microbiologia, BIOAGRO, Universidade Federal de Viçosa, Viçosa, MG, Brazil |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Colombo |D Lívia |u Laboratório de Fisiologia de Micro-organismos, Departamento de Microbiologia, BIOAGRO, Universidade Federal de Viçosa, Viçosa, MG, Brazil |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Roberti |D Alvaro |u Laboratório de Fisiologia de Micro-organismos, Departamento de Microbiologia, BIOAGRO, Universidade Federal de Viçosa, Viçosa, MG, Brazil |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Alvim |D Mariana |u Laboratório de Fisiologia de Micro-organismos, Departamento de Microbiologia, BIOAGRO, Universidade Federal de Viçosa, Viçosa, MG, Brazil |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Cardoso |D Silvia |u Laboratório de Imunovirologia Molecular, Departamento de Biologia Geral, Universidade Federal de Viçosa, Viçosa, MG, Brazil |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Reis |D Kledna |u Microbiologia Veterinária Especial - MICROVET, Viçosa, MG, Brazil |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a de Paula |D Sérgio |u Laboratório de Imunovirologia Molecular, Departamento de Biologia Geral, Universidade Federal de Viçosa, Viçosa, MG, Brazil |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a da Silveira |D Wendel |u Laboratório de Fisiologia de Micro-organismos, Departamento de Microbiologia, BIOAGRO, Universidade Federal de Viçosa, Viçosa, MG, Brazil |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Passos |D Flavia |u Laboratório de Fisiologia de Micro-organismos, Departamento de Microbiologia, BIOAGRO, Universidade Federal de Viçosa, Viçosa, MG, Brazil |4 aut | ||
| 950 | |B NATIONALLICENCE |P 773 |E 0- |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/3(2015-02-01), 1191-1203 |x 0175-7598 |q 99:3<1191 |1 2015 |2 99 |o 253 | ||