XXII. BIOCHEMICKÝ ZJAZD - Jesseniova lekárska fakulta

Posters
11.
DESIGN of an EXPrESSION SYSTEM for THE PRODUCTION OF
RECOMBINANT HUMAN THROMBIN IN ESCherICHIA COLI
Michaela Kandričáková 1 , Stanislav Stuchlík1 and Ján Turňa 1,2
1
Comenius University, Faculty of Natural Sciences, Department of Molecular Biology,
2
Department of Molecular Biology SAV Bratislava
Thrombin is a plasmatic protease, which plays a key role in blood coagulation. This form of
active prothrombin is characterised by many biological functions including procoagulation
and anti coagulation effects. The polyfunctionality of thrombin has received increasing
interest in biofarmaceutical and biomedical research. One of the possible ways to gain
recombinant human thrombin is to use expression systems based on E.coli cells. E.coli
offers many adventages over other host organisms and therefore remains the most
commonly used host for the production of heterologous proteins. Prethrombin-2, an
activation intermediate of thrombin, is often expressed as a precursor of prothrombin.
Presumably, the use of this gene assures higher transcription and translation efficiency
compared to prothrombin gene mainly because of the smaller size of the mRNA transcript.
During the preparation of prethrombin-2 syntetic gene the known sequence will be used
with optimized E.coli codon usage ensuring higher expression yields.
The aim of this work is the design of proper expression system for the production of
recombinant human thrombin in the E.coli host cells. The study is focused on selection of
proper gene sequence with codon usage optimized for E.coli, selection of suitable expression
system and in the next step we would like to optimise physiological conditions for the
foreign protein expression and subsequent verify expression of recombinant thrombin.
126 XXII. Biochemistry Congress, Martin