Nuclear Import Analysis of Two Different Fluorescent Marker Proteins into Hepatocyte Cell Lines (HuH-7 Cell)

• Main Authors: Aris Haryanto, Michael Kann
• Format: Article
• Language: English
• Published: Universitas Gadjah Mada, Yogyakarta 2015-10-01
• Series: Indonesian Journal of Biotechnology
• Online Access: http://journal.ugm.ac.id/ijbiotech/article/view/7557

The application of fluorescent proteins as expression markers and protein fusion partners has proved<br />immensely valuable for resolving the organization of biological events in living cells. EGFP and DsRed2 are<br />commonly fluorescent marker protein which is used for biotechnology and cell biology research. The present<br />study was designed to identify the expression vector that suitable to ligate with DNA encoding HBV core<br />protein for intracellular localization study in hepatocyte cell, which were expressed as fusion proteins. We also<br />compared and quantified the expressed fluorescent protein which predominantly localized in the cell<br />compartment. The results indicated that DsRed2 shown as less than ideal for intracellular localization study of<br />than EGFP, because of its tetrameric structure of the fluorescent protein and when fused to a protein of interest,<br />the fusion protein often forms aggregates in the living cells. In contrast, EGFP fluorescent protein shown a much<br />higher proportion of cytoplasmic localization, thus being more suitable for analysis of intracellular localization<br />than DsRed2 fluorescent protein. EGFP fluorescent protein is also capable to produce a strong green fluorescence<br />when excited by blue light, without any exogenously added substrate or cofactor, events inside living cell can<br />thus be visualized in a non-invasive way. Based on our present quantitative data and some reasons above shown<br />that EGFP is more suitable than DsRed2 as a fluorescent marker protein for intracellular localization study into<br />HuH-7 cell.<br />Keywords: EGFP, DsRed2 fluorescent protein , HuH-7 cell, HBV, intracellular localization