k 2016

Monogenea: parasite-host interactions at molecular level

KAŠNÝ, Martin, Jana ILGOVÁ, Jiří VOREL, Pavel ROUDNICKÝ, Libor MIKEŠ et. al.

Základní údaje

Originální název

Monogenea: parasite-host interactions at molecular level

Autoři

KAŠNÝ, Martin (203 Česká republika, domácí), Jana ILGOVÁ (203 Česká republika), Jiří VOREL (703 Slovensko), Pavel ROUDNICKÝ (203 Česká republika), Libor MIKEŠ (203 Česká republika), Hana DVOŘÁKOVÁ (203 Česká republika), Lucie JEDLIČKOVÁ (203 Česká republika), Dagmar JIRSOVÁ (203 Česká republika), Hynek STRNAD (203 Česká republika), Roman LEONTOVYČ (203 Česká republika), Ewa DZIKA (203 Česká republika), Božena KOUBKOVÁ (203 Česká republika), Lukáš VETEŠNÍK (203 Česká republika), Pavel JURAJDA (203 Česká republika) a Milan GELNAR (203 Česká republika, garant)

Vydání

5th ECIP meeting - European Centre of Ichtyoparasitology, 2016

Další údaje

Jazyk

angličtina

Typ výsledku

Prezentace na konferencích

Obor

10600 1.6 Biological sciences

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Kód RIV

RIV/00216224:14310/16:00088397

Organizační jednotka

Přírodovědecká fakulta

ISBN

978-80-210-8373-8

Klíčová slova anglicky

Monogenea; Eudiplozoon; nipponicum; hematophagy; inhibitors; peptidases
Změněno: 25. 3. 2017 08:03, RNDr. Martin Kašný, Ph.D.

Anotace

V originále

In a first part of our pilot study of monogeneans molecules we adopted the NGS techniques in order to get the high quality “monogenean-polyopisthocotylean genome/transcriptome matrix”. In a second part of our study this platform was used for the identification of the monogeneans´ dominant protein molecules followed by their further molecular/biochemical characterization. For this purpose we adopted Eudiplozoon nipponicum as a experimental model species; the representative of blood-feeding monogeneans (family Diplozoidae) of fresh water fishes, the ectoparasite from the gills of Cyprinus carpio. The bioinformatic analyses of genomic sequence data were performed; the first genome assembly of 98 mil of E. nipponicum DNA (3 libraries) sequence reads was done and the calculation of genome size was initiated. Up to now, the low coverage (6%) was reached and therefore the distorted number of bases of whole E. nipponicum genome (400 Gb) was estimated. By adoption of homology searches of RNA 158 753 271 bases/223 887 transcripts the 9757 contigs (>1 kb) were assembled and particular protein molecules in E. nipponicum transcriptome data were identified, e.g. peptidases/inhibitors; 29 contigs of cysteine peptidases (e.g. cathepsin L) and 7 contigs of their inhibitors (e.g. cystatins); 12 contigs of serine peptidases (e.g. cathepsin A) and 7 contigs of their inhibitors (e.g. serpin). Employing biochemical, proteomic and molecular tools, we found that cysteine peptidase activities prevailed in soluble protein extracts and excretory/secretory (E/S) products of E. nipponicum and the major part of activity was related to cathepsin L-like. Mass spectrometry revealed several tryptic peptides in E/S products matching to two translated sequences of cathepsin L genes. The dominance of cysteine peptidases of cathepsin L type in E. nipponicum resembles the situation in, e.g., fasciolid trematodes. The cathepsin L3 was cloned and expressed in both bacterial and yeasts expression systems. The recombinant enzyme was purified on Ni-NTA agarose column and the experiments focused on its molecular/biochemical properties were started.

Návaznosti

GBP505/12/G112, projekt VaV
Název: ECIP - Evropské centrum ichtyoparazitologie
Investor: Grantová agentura ČR, ECIP - Evropské centrum ichtyoparazitologie
Zobrazeno: 19. 11. 2024 09:59