Detailed Information on Publication Record
2015
Expression of a cysteine peptidase inhibitor from Eudiplozoon nipponicum (Monogenea).
ILGOVÁ, Jana, Milan GELNAR and Martin KAŠNÝBasic information
Original name
Expression of a cysteine peptidase inhibitor from Eudiplozoon nipponicum (Monogenea).
Authors
ILGOVÁ, Jana (703 Slovakia, guarantor, belonging to the institution), Milan GELNAR (203 Czech Republic, belonging to the institution) and Martin KAŠNÝ (203 Czech Republic, belonging to the institution)
Edition
22nd Helminthological Days, 2015
Other information
Language
English
Type of outcome
Konferenční abstrakt
Field of Study
10600 1.6 Biological sciences
Country of publisher
Czech Republic
Confidentiality degree
není předmětem státního či obchodního tajemství
RIV identification code
RIV/00216224:14310/15:00080924
Organization unit
Faculty of Science
ISBN
978-80-7444-032-8
Keywords in English
cystatin; recombinant protein expression; Eudiplozoon nipponicum
Změněno: 24/7/2015 10:11, Mgr. et Mgr. Jana Ilgová, Ph.D.
Abstract
V originále
Our research is focused primarily on expression and characterization of cystatin produced by platyhelminth Eudiplozoon nipponicum. This blood-feeding ectoparasite of Cyprinus carpio (common carp) is a representative of the family Diplozooidae (Monogenea). Cystatins – cysteine peptidase inhibitors are produced by a wide range of organisms and belong to bioactive molecules with immunomodulatory and inhibitory properties. The transcriptomic data of E. nipponicum were analyzed for the presence of cystatin sequences. The partial nucleotide sequence was identified and the whole gene sequence was obtained by PCR, 3’/5’ RACE PCR and sequencing using cDNA as a template. Gene coding 98 amino acid cystatin of E. nipponicum with predicted molecular weight 10.85 kDa and theoretical pI 6.27 was inserted into pET19b expression vector and the obtained construct was transferred into E. coli competent cells (BL 21 strain). Expression of recombinant cystatin molecule was induced by adding IPTG to the cultivation media. The success of production was confirmed by mass spectrometry in a dominant protein band after the fractionation of the E. coli derived protein mixture by SDS-PAGE. Recombinant protein was produced in the insoluble form incorporated into inclusion bodies. The cystatin from E. nipponicum will be subsequently solubilized and tested for its functional and structural properties.
Links
| GAP506/12/1258, research and development project |
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| GBP505/12/G112, research and development project |
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| MUNI/A/1484/2014, interní kód MU |
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