Expression of non-structural proteins of Potato mop-top virus in procaryotic system

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60460709%3A41210%2F04%3A8778" target="_blank" >RIV/60460709:41210/04:8778 - isvavai.cz</a>

Result on the web

—

DOI - Digital Object Identifier

—

Result language

angličtina

Original language name

Expression of non-structural proteins of Potato mop-top virus in procaryotic system

Original language description

PMTV RNA 1 was divided into 6 parts with the length 1 kb aproximately, and these fragments were inserted into pUC-57A suitable for sequencing. The obtained sequences showed high homology (GenBank AY196959). All fragments were inserted into different expression vectors. First of all we used the non-tagged expression vectors, PMPM-&#937;-4, 5, 6; pET-21a+ and pTH. To facilitate detection of expressed proteins we used tagged vectors - either with His tag or glutation S-transferase (GST). GST fusion proteinwas performed in vectors pGEX-4T-1, 2 and pGEX-KG alowing the removing of tag by thrombin. Proteins with His tag were induced in vectors pQE-40 and pQE-60. The highest yield of expressed protein was obtained from pGEX-4T-2 in E. coli BL 21. Disadvantageof this aproach was uncomplete cleavage of GST by thrombin. We consider pQE-40 and pQE-60 as more suitable vectors for the purpose of production of polyclonal antibodies against PMTV proteins, because sequence of tag 6xHis has not been f

Czech name

Exprese nestrukturálních proteinů mop-top viru bramboru v bakteriálním systému

Czech description

PMTV RNA 1 was divided into 6 parts with the length 1 kb aproximately, and these fragments were inserted into pUC-57A suitable for sequencing. The obtained sequences showed high homology (GenBank AY196959). All fragments were inserted into different expression vectors. First of all we used the non-tagged expression vectors, PMPM-&#937;-4, 5, 6; pET-21a+ and pTH. To facilitate detection of expressed proteins we used tagged vectors - either with His tag or glutation S-transferase (GST). GST fusion proteinwas performed in vectors pGEX-4T-1, 2 and pGEX-KG alowing the removing of tag by thrombin. Proteins with His tag were induced in vectors pQE-40 and pQE-60. The highest yield of expressed protein was obtained from pGEX-4T-2 in E. coli BL 21. Disadvantageof this aproach was uncomplete cleavage of GST by thrombin. We consider pQE-40 and pQE-60 as more suitable vectors for the purpose of production of polyclonal antibodies against PMTV proteins, because sequence of tag 6xHis has not been f

Type

D - Article in proceedings

CEP classification

GF - Diseases, pests, weeds and plant protection

OECD FORD branch

—

Project

—

Continuities

Z - Vyzkumny zamer (s odkazem do CEZ)

Publication year

2004

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Article name in the collection

Proceeding of the XVI. Slovak and Czech Plant Protection Conference

ISBN

1335-258X

ISSN

—

e-ISSN

—

Number of pages

3

Pages from-to

257-259

Publisher name

Slovak Agricultural Universitty in Nitra

Place of publication

Nitra

Event location

Nitra

Event date

Sep 16, 2003

Type of event by nationality

WRD - Celosvětová akce

UT code for WoS article

—