| Abstract [eng] |
The aim of this work was to investigate the recombinant mink growth hormone (mGH) inclusion bodies composition, formed in E. coli induced with different concentrations of inductor. Unknown proteins quantity in insoluble fraction was compared between cultures induced with different inductor concentrations. Different concentrations of inductor was used: 500 µM (control), 50 µM and 10 µM. Cell cultivation was performed at 25 ⁰C degree and samples were collected 27 hours after induction. To assess quantitative differences between samples 2-D SDS-PAGE electrophoresis method was used, gels were processed with Progenesis SameSpots program and results were statistically analyzed. Proteins of interest were analyzed with MS/MS mass spectrometer Orbitrap Velos coupled with UPLC Waters nanoAcquity chromatograph. Proteins were identified with MASCOT program in Sprot database. Six proteins were identified. Higher amount of chaperone DnaK and enzyme beta-lactamase TEM was detected in samples with low induction. Lower amount of periplasmic oligopeptide-binding protein was detected in samples with low level of induction compared with control samples. Structure: introduction, part of theory, experimental part, results and discussion, conclusions, references. Thesis consist of: 56 p., 7 pictures, 5 tables, 66 bibliographical entries. |
