| dc.description.abstract | Heparin-binding EGF-like growth factor (HB-EGF) is a member of the EGF protein
family characterized by specific growth stimulation of many cells with important
physiological functions. HB-EGF is shown to have many pivotal applications in
researches, clinics and therapeutics; therefore, the needs of HB-EGF for health care
in Vietnam is quite important. However, the production of HB-EGF in standard
approach of E.Coli encountered various problems including low yield, aggregation,
besides, its high cost and short of supply. My thesis studies aim to employ a better
expression vector for HB-EGF production. If successful, this work could facilitate the
production of high yield and functional protein. We also hope the expressed proteins
could be used for work at the Stem Cell Laboratory such as tissues culture, analytical
assays and other experimental studies.
In our study, the HB-EGF gene is synthesized by one step reverse transcription
polymerase chain reaction (RT-PCR) using total RNA of human cord bloods. After the
gene of interest is obtained, TA cloning is employed to clone the HB-EGF gene into
the pET SUMO expression vector. The candidate clones were screened by polymerase
chain reaction (PCR) and selected for structural analysis by sequencing. The result
showed the identical between the selected genes and the HB-EGF gene sequence of
references in NCBI. For future studies, the clones will be expressed in a suitable host
to produce HB-EGF protein.
Key word: HB-EGF, TA cloning, RT-PCR, PCR, SUMO expression vector. | en_US |
