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UTVRĐIVANJE USPEŠNOSTI TRANSFEKCIJE I EKSPRESIJE SOX14 U HELA
ĆELIJAMA

THE ANALYSIS OF TRANSFECTION EFFICIENCY AND EXPRESSION OF SOX14 IN
HELA CELLS

Autor: MANOJLOVID MILICA Treda beogradska gimnazija – I razred Regionalni centar za talente Beograd 2

Mentori: dr ANDRIJANA KLAJN, naučni saradnik, Institut za molekularnu genetiku i genetičko inženjerstvo,
Univerzitet u Beogradu DANIJELA STANISAVLJEVID, istraživač - saradnik Institut za molekularnu genetiku i genetičko
inženjerstvo, Univerzitet u Beogradu Student-mentor: STEFAN LAZID,

REZIME
Iako je humani SOX14 gen kloniran i okarakterisan pre više od jedne decenije, malo se zna o molekularnim
mehanizmima i delijskim procesima u koje je uključen. U cilju razjašnjavanja uloge SOX14 u različitim fiziološkim i
patološkim procesima, kodirajuda sekvenca humanog SOX14 gena uklonirana je u ekspresioni pcDNA3.1 vektor. Cilj
ovog rada je bio utvrđivanje efikasnosti transfekcije i ekspresije humanog SOX14 gena u HeLa delijama. HeLa delije
su kotransfektovane sa pcDNASOX14 konstruktom i sa pEGFP-C1 vektorom sa kojeg se eksprimira zeleni
florescentni protein (GFP). Na osnovu broja GFP-pozitivnih delija ustanovljeno je da je 16% HeLa delija primilo
konstrukte. Western blot metodom pokazano je da je i pored relativno niskog nivoa transfekcija, nivo ukupnog
SOX14 proteina u uzorku transfektovanih delija značajno vedi u odnosu na kontrole (HeLa delije bez transfekcije i
HeLa transfektovane pcDNA3.1 praznim vektorom). Na osnovu dobijenih rezultata može se zaključiti da strategija,
pod opisanim uslovima u ovom radu, može biti primenjena za planirana dalja istraživanja koja uključuju i
proučavanje potencijalne antikancerogene aktivnosti SOX14 proteina preko apoptoznog efekta u HeLa delijama.

KLJUČNE REČI: Transfekcija, Western blot, HeLa delije, SOX14

SUMMARY

Although the human SOX14 gene was first cloned and characterized more than a decade ago, little is know about
its involvement in different molecular mechanisms and cellular processes. In order to elucidate the role of SOX14 in
diverse physiological and pathophysiological processes, coding sequence of human SOX14 gene was cloned in
pcDNA3.1 expression vector. The aim of this work was to determine the transfection efficiency and expression of
human SOX14 gene in HeLa cells. These cells were co-transfected with pcDNASOX14 construct and pEGFP-C1 vector
containing the sequence coding for Green Florescent Protein (GFP). According to the number of GFP-positive cells,
16% of HeLa cells were successfully co-transfected. In spite of relatively low transfection efficiency, the overall
SOX14 protein level was significantly higher in the sample containing transfected cells comparing to the controls
(HeLa cells without transfection and HeLa cells transfected with pcDNA3.1 empty vector) as revealed by Western
Blot. In accordance with the obtained results, we can conclude that the proposed strategy, under conditions
described in this work, can be applied in future research including the analyses of potential anti-cancer activity of
SOX14 via apoptotic effect in HeLa cells.

KEY WORDS: Transfection, Western blot HeLa cells, SOX14