Controlled expression of enhanced green fluorescent protein and hepatitis b virus precore protein in mammalian cells

2003 ◽  
Vol 13 (2) ◽  
pp. 114
Author(s):  
Xiaodong ZHU
Keyword(s):  
Green Fluorescent Protein ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Mammalian Cells ◽  
Enhanced Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
B Virus ◽  
Green Fluorescent

scholarly journals Functional incorporation of green fluorescent protein into hepatitis B virus envelope particles

Virology ◽  
2004 ◽  
Vol 330 (1) ◽  
pp. 158-167 ◽  
Author(s):  
Carsten Lambert ◽  
Nicole Thomé ◽  
Christoph J. Kluck ◽  
Reinhild Prange
Keyword(s):  
Green Fluorescent Protein ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Fluorescent Protein ◽  
Virus Envelope ◽  
B Virus ◽  
Green Fluorescent

scholarly journals High-Titer Retroviral Vectors Containing the Enhanced Green Fluorescent Protein Gene for Efficient Expression in Hematopoietic Cells

Blood ◽  
1997 ◽  
Vol 90 (9) ◽  
pp. 3316-3321 ◽  
Author(s):  
Ana Limón ◽  
Javier Briones ◽  
Teresa Puig ◽  
Mercé Carmona ◽  
Oscar Fornas ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Green Fluorescent Protein ◽  
Mammalian Cells ◽  
Enhanced Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Hematopoietic Cells ◽  
Retroviral Vectors ◽  
Green Fluorescent Protein Gene ◽  
Efficient System ◽  
Green Fluorescent

Abstract Retroviral vectors constitute the most efficient system to deliver and integrate foreign genes into mammalian cells. We have developed a producer cell line that yields high titers of amphotropic retroviral vectors carrying the enhanced green fluorescent protein (EGFP) gene, a codon humanized, red-shifted variant of the green fluorescent protein (GFP) gene, which can be used as a selectable marker. We have used a hybrid vector that has been shown to efficiently drive gene expression in hematopoietic cells. Virtually all murine and human cell lines and primary human hematopoietic cells tested were transduced with varying efficiency after incubation with vector-containing supernatants. Human CD34+ cells obtained from cord blood or aphereses products were transduced using a protocol that involves daily addition of vector-containing supernatants for 6 consecutive days. At day 6, up to 16% of the cells expressed EGFP, as assessed by flow cytometry. Sorted EGFP-expressing cells were able to produce fluorescent hematopoietic colonies. EGFP's main advantages are its fast flow cytometry determination and the possibility of cell sorting and simultaneous evaluation of the transduction efficiency along with other phenotypic markers.

scholarly journals Formation of Vesicular Stomatitis Virus Pseudotypes Bearing Surface Proteins of Hepatitis B Virus

2005 ◽  
Vol 79 (19) ◽  
pp. 12566-12574 ◽  
Author(s):  
Manujendra N. Saha ◽  
Atsushi Tanaka ◽  
Atsushi Jinno-Oue ◽  
Nobuaki Shimizu ◽  
Kazushi Tamura ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Cell Lines ◽  
Vesicular Stomatitis Virus ◽  
Fluorescent Protein ◽  
Surface Proteins ◽  
B Virus ◽  
Vesicular Stomatitis ◽  
Green Fluorescent ◽  
Short Time

ABSTRACT It has been difficult to propagate and titrate hepatitis B virus (HBV) in tissue culture. We examined whether vesicular stomatitis virus (VSV) pseudotypes bearing HBV surface (HBs) proteins infectious for human cell lines could be prepared. For this, expression plasmids for three surface proteins, L, M, and S, of HBV were made. 293T cells were then transfected with these plasmids either individually or in different combinations. 293T cells expressing HBs proteins were infected with VSVΔG*-G, a recombinant VSV expressing green fluorescent protein (GFP), to make VSV pseudotypes. Culture supernatants together with cells were harvested and sonicated for a short time. The infectivities of freshly harvested supernatants were determined by quantifying the number of cells expressing GFP after neutralization with anti-VSV serum and mouse monoclonal antibodies (MAbs) against HBs protein. Among 14 cell lines tested for susceptibility to HBV pseudotype samples, HepG2, JHH-7, and 293T cells were judged to be the most susceptible. Namely, the infectious units (IU) of the culture supernatant samples neutralized with anti-VSV in the absence and presence of anti-HBs S MAbs and titrated on HepG2 cells ranged from 1,000 to 4,000 IU/ml and 200 to 400 IU/ml, respectively, suggesting the presence of VSVΔG*(HBV) pseudotypes. This infectivity was inhibited by treatment with lactoferrin or dextran sulfate. Pretreatment of the cells with trypsin or tunicamycin inhibited plating of the pseudotype samples. The HBV pseudotypes can be used to analyze early steps of HBV infection, including the entry mechanism of HBV.

scholarly journals Enhanced Green Fluorescent Protein (GFP) fluorescence after polyelectrolyte caging

2006 ◽  
Vol 14 (21) ◽  
pp. 9815 ◽  
Author(s):  
Alberto Diaspro ◽  
Silke Krol ◽  
Barbara Campanini ◽  
Fabio Cannone ◽  
Giuseppe Chirico
Keyword(s):  
Green Fluorescent Protein ◽  
Enhanced Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Gfp Fluorescence ◽  
Green Fluorescent
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