High-level secretion of functional green fluorescent protein from transgenic tobacco cell cultures: Characterization and sensing

2004 ◽  
Vol 85 (6) ◽  
pp. 610-619 ◽  
Author(s):  
Wei Wen Su ◽  
Peizhu Guan ◽  
Robert C. Bugos
Keyword(s):  
Green Fluorescent Protein ◽  
Transgenic Tobacco ◽  
Cell Cultures ◽  
Fluorescent Protein ◽  
Tobacco Cell ◽  
Green Fluorescent ◽  
High Level

High level Purkinje cell specific expression of green fluorescent protein in transgenic mice

2001 ◽  
Vol 115 (6) ◽  
pp. 455-464 ◽  
Author(s):  
Xulun Zhang ◽  
Stephan L. Baader ◽  
Feng Bian ◽  
Wolfgang Müller ◽  
John Oberdick
Keyword(s):  
Green Fluorescent Protein ◽  
Transgenic Mice ◽  
Purkinje Cell ◽  
Fluorescent Protein ◽  
Specific Expression ◽  
Green Fluorescent ◽  
High Level

scholarly journals In Vitro Replication of Hepatitis E Virus (HEV) Genomes and of an HEV Replicon Expressing Green Fluorescent Protein

2004 ◽  
Vol 78 (9) ◽  
pp. 4838-4846 ◽  
Author(s):  
Suzanne U. Emerson ◽  
Hanh Nguyen ◽  
Judith Graff ◽  
David A. Stephany ◽  
Alicia Brockington ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Cell Cultures ◽  
Hepatitis E Virus ◽  
Fluorescent Protein ◽  
Hepatitis E ◽  
Green Fluorescent Protein Gene ◽  
Primate Cell ◽  
Green Fluorescent ◽  
Full Length Genome

ABSTRACT Hepatitis E virus (HEV) RNA replication occurred in seven of nine primate cell cultures transfected with in vitro transcripts of an infectious cDNA clone. Cell-to-cell spread did not occur in cell cultures, but rhesus monkeys inoculated with lysates of HEV-transfected PLC/PRF/5 and Huh-7 cells became infected with HEV. A replicon with the ORF2 and ORF3 genes deleted and replaced with the green fluorescent protein gene also replicated in the same primate cells that supported the replication of the full-length genome. Fluorescence-activated cell sorter analysis confirmed that the 7mG cap structure was critical for efficient infectivity, although replication could be initiated at a very low level in its absence. HEV virions were also able to infect a limited number of cells of certain lines.

scholarly journals Multicopy Integration and Expression of Heterologous Genes in Methylobacterium extorquens ATCC 55366

2006 ◽  
Vol 72 (1) ◽  
pp. 753-759 ◽  
Author(s):  
Young J. Choi ◽  
Denis Bourque ◽  
Lyne Morel ◽  
Denis Groleau ◽  
Carlos B. Míguez
Keyword(s):  
Green Fluorescent Protein ◽  
Target Genes ◽  
Fluorescent Protein ◽  
Expression System ◽  
Attachment Site ◽  
Methylobacterium Extorquens ◽  
Gene Copies ◽  
Multicopy Integration ◽  
Green Fluorescent ◽  
High Level

ABSTRACT High-level expression of chromosomally integrated genes in Methylobacterium extorquens ATCC 55366 was achieved under the control of the strong M. extorquens AM1 methanol dehydrogenase promoter (P mxaF ) using the mini-Tn7 transposon system. Stable maintenance and expression of the integrated genes were obtained in the absence of antibiotic selective pressure. Furthermore, using this technology, a multicopy integration protocol for M. extorquens was also developed. Chromosomal integration of one to five copies of the gene encoding the green fluorescent protein (gfp) was achieved. The multicopy-based expression system permitted expression of a preset number of gene copies. A unique specific Tn7 integration locus in the chromosome of M. extorquens, known as the Tn7 attachment site (attTn7 site), was identified. This single attTn7 site was identified in an intergenic region between glmS, which encodes the essential enzyme glucosamine-6-phosphate synthetase, and dhaT, which encodes 1,3-propanediol dehydrogenase. The fact that the integration event is site specific and the fact that the attTn7 site is a noncoding region of the chromosome make the mini-Tn7 transposon system very useful for insertion of target genes and subsequent expression. In all transformants tested, expression and segregation of the transforming gene were stable without generation of secondary mutations in the host. In this paper, we describe single and multicopy chromosome integration and stable expression of heterologous genes (bgl [β-galactosidase], est [esterase], and gfp [green fluorescent protein]) in M. extorquens.

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