[Effects of signal peptide sequence of hemagglutinin on the expression of green fluorescent protein and hemagglutinin fusion gene].
OBJECTIVE
Purified avian influenza virus (AIV) hemagglutinin (HA) gene fragment was inserted into green fluorescent protein (GFP) expression vector, pEGFP-C1. The role of signal peptide in the HA-GFP expression in 293T cells was investigated by cutting the signal peptide or placing it in different locations of HA-GFP fusion gene.


METHODS
The expression of GFP in 293T cells was examined directly with fluorescence microscope and flow cytometry analysis.


RESULTS
After transfected with pEGFP-C1, M1, M2 and M3 plasmid, under fluorescent microscope the HA-GFP fusion protein was successfully expressed. Fluorescence was seen homogeneously distributed in the entire cell body of the cells transfected by the empty vector pEGFP-C1, whereas signal peptide could obviously reduced the expression of GFP-HA compared with recombinant plasmid without signal peptide. In addition, we found that there was no significant effect of the location of signal peptide on the expression of HA-GFP fusion protein.
Su
Yan
Y
College of Animal Medicine, Xinjiang Agricultural University, Urumqi 830052, China. 2006au@163.com
Zhang
Baojiang
B
Shen
Yu
Y
Huang
Jiasi
J
D002460:Cell Line; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D006388:Hemagglutinins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D009980:Influenza A virus / Q000378:metabolism; D008856:Microscopy, Fluorescence; D016133:Polymerase Chain Reaction; D021382:Protein Sorting Signals / Q000235:genetics / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*
D006388:Hemagglutinins; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins