scholarly journals Generation of a Stable Transgenic Swine Model Expressing a Porcine Histone 2B-eGFP Fusion Protein for Cell Tracking and Chromosome Dynamics Studies

PLoS ONE ◽  
2017 ◽  
Vol 12 (1) ◽  
pp. e0169242 ◽  
Author(s):  
Renan B. Sper ◽  
Sehwon Koh ◽  
Xia Zhang ◽  
Sean Simpson ◽  
Bruce Collins ◽  
...  
FEBS Letters ◽  
2003 ◽  
Vol 542 (1-3) ◽  
pp. 142-146 ◽  
Author(s):  
Shengnan Jin ◽  
Timothy D McKee ◽  
Daniel D Oprian

2001 ◽  
Vol 29 (3) ◽  
pp. A82-A82
Author(s):  
K. Pattni ◽  
M. Jepson ◽  
H. Stenmark ◽  
G. Banting

2002 ◽  
Vol 7 (4) ◽  
pp. 325-332 ◽  
Author(s):  
Arno Pol ◽  
Fred Van Ruissen ◽  
Joost Schalkwijk

Inflamed epidermis (psoriasis, wound healing, ultraviolet-irradiated skin) harbors keratinocytes that are hyperproliferative and display an abnormal differentiation program. A distinct feature of this so-called regenerative maturation pathway is the expression of proteins such as the cytokeratins CK6, CK16, and CK17 and the antiinflammatory protein SKALP/elafin. These proteins are absent in normal skin but highly induced in lesional psoriatic skin. Expression of these genes can be used as a surrogate marker for psoriasis in drug-screening procedures of large compound libraries. The aim of this study was to develop a keratinocyte cell line that contained a reporter gene under the control of a psoriasis-associated endogenous promoter and demonstrate its use in an assay suitable for screening. We generated a stably transfected keratinocyte cell line that expresses enhanced green fluorescent protein (EGFP), under the control of a 0.8-kb fragment derived from the promoter of the SKALP/elafin gene, which confers high levels of tissue-specific expression at the mRNA level. Induction of the SKALP promoter by tumor necrosis factor-ca resulted in increased expression levels of the secreted SKALP-EGFP fusion protein as assessed by direct readout of fluorescence and fluorescence polarization in 96-well cell culture plates. The fold stimulation of the reporter gene was comparable to that of the endogenous SKALP gene as assessed by enzyme-linked immunosorbent assay. Although the dynamic range of the screening system is limited, the small standard deviation yields a Z factor of 0.49. This indicates that the assay is suitable as a high-throughput screen, and provides proof of the concept that a secreted EGFP fusion protein under the control of a physiologically relevant endogenous promoter can be used as a fluorescence-based high-throughput screen for differentiation-modifying or antiinflammatory compounds that act via the keratinocyte.


2008 ◽  
Vol 30 (8) ◽  
pp. 1409-1414 ◽  
Author(s):  
Jin-bao Tang ◽  
Peng Zhu ◽  
Hong-ming Yang ◽  
Li-min Sun ◽  
Shu-liang Song ◽  
...  

2015 ◽  
Vol 31 (3) ◽  
pp. 641-649 ◽  
Author(s):  
Hsing Chieh Wu ◽  
Yu San Chen ◽  
Pin Chun Shen ◽  
Jui Hung Shien ◽  
Long Huw Lee ◽  
...  

2004 ◽  
Vol 72 (5) ◽  
pp. 2810-2816 ◽  
Author(s):  
Camilo Cuadros ◽  
Francisco J. Lopez-Hernandez ◽  
Ana Lucia Dominguez ◽  
Michael McClelland ◽  
Joseph Lustgarten

ABSTRACT Vaccination is the most efficient prophylaxis against a variety of infectious diseases. New vaccination strategies rely on the incorporation of effective adjuvants, which stimulate the innate immune response and, in turn, activate the adaptive immune response. It is well established that flagellin induces inflammatory responses through the activation of antigen-presenting cells (APCs). In order to evaluate whether flagellin can serve as a carrier for the development of adjuvants or vaccines, we prepared a flagellin-enhanced green fluorescent protein (EGFP) fusion protein. Our results demonstrate that a flagellin-EGFP fusion protein is capable of stimulating APCs, resulting in the maturation of these cells and secretion of proinflammatory cytokines. Furthermore, APCs pulsed with the flagellin-EGFP fusion protein effectively process and present EGFP antigens. More importantly, animals immunized with the flagellin-EGFP fusion protein developed specific anti-EGFP T-cell responses. In contrast, recombinant EGFP was not able to stimulate APCs, nor did it induce a T-cell response. Thus, recombinant-flagellin fusion proteins may be suitable carriers as adjuvants or vaccines for the development of new vaccination strategies to induce and boost immune responses against infectious diseases and cancer.


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