Klonování mannosyltransferas z Aspergillus fumigatus

Mgr. Andrea Fleková

Bachelor's thesis

Cloning of mannosyltransferases from Aspergillus fumigatus

Abstract:

Tato práce se věnuje studiu mannosyltransfras se zaměřením na β-1,4-mannosyltransferasu z Aspergillus fumigatus. Mannosyltransferasy hrají u A. fumigata významnou roli ve stavbě buněčné stěny. Protože A. fumigatus způsobuje onemocnění zvané aspergilosa u imunodeficitních jedinců, je buněčná stěna v neustálém kontaktu s hostitelským organismem a hraje hlavní roli v patogenitě a virulenci. Poruchy mannosylace u tohoto patogena vedou k oslabení buněčné stěny a nižší odolnosti vůči prostředí. Inhibice funkce mannosyltransferas může být využita jako terapeutický cíl. β-1,4-mannosyltransferasa byla vyhledána pomocí databáze CAZy a bylo zjištěno, že tato mannosyltransferasa obsahuje 505 aminokyselin. Gen pro tento protein byl úspěšně vložen do vektoru pET29b a ten byl úspěšně transformován do expresních buněk E. coli. Tyto expresní buňky budou následně použity pro expresi proteinu.

Abstract:

This work is focused on studying of mannosyltransferases, specifically β-1,4-mannosyltransferase from Aspergillus fumigatus. Mannosyltransferases play an essential role in construction of the cell wall in A. fumigatus. Because A. fumigatus causes the disease called Aspergillosis by immunosuppresion individuals, the cell wall is in continuous contact with host organism and play major role in pathogenicity and virulence. Disorders of mannosylation by this pathogen leads to weakening of the cell wall and less resistance to the environment. The inhibition of the function of mannosyltransferases can be used as the therapeutic target. β-1,4-mannosyltransferase was found in the database CAZy and it was shown, that this mannosyltransferase contained 505 amino acid. Gene for this protein was successfully cloned in the vector pET29b and the resulting plasmid was successfully transformed into E. coli express cells. The cells are ready for subsequent expression of the protein.

Keywords

Klonování, mannosyltransferasy, Aspergillus fumigatus

Language used: Czech

Date on which the thesis was submitted / produced: 6. 5. 2009

Thesis defence

• Date of defence: 16. 6. 2009
• Supervisor: prof. RNDr. Michaela Wimmerová, Ph.D.

Masaryk University

Faculty of Science

Bachelor programme / field:
Biochemistry / Biochemistry