scholarly journals Identification of genes differentially regulated by glucocorticoids and progestins using a Cre/loxP-mediated retroviral promoter-trapping strategy

2002 ◽  
Vol 28 (3) ◽  
pp. 177-192 ◽  
Author(s):  
Y Wan ◽  
SK Nordeen
Keyword(s):  
Target Genes ◽  
Selectable Marker ◽  
Biological Effects ◽  
Fibroblast Cell ◽  
Single Copy ◽  
Mouse Fibroblast ◽  
Marker Genes ◽  
Flanking Sequence ◽  
Selectable Marker Genes ◽  
Promoter Trap

Glucocorticoids and progestins are two classes of steroid hormone with very distinct biological functions. However, the glucocorticoid receptor (GR) and the progesterone receptor (PR) share many structural and functional similarities. One way that glucocorticoids and progestins can exert different biological effects is through their different abilities to regulate the expression of certain target genes. A strategy employing a retroviral promoter-trap and Cre/loxP-mediated site-specific recombination has been developed to identify genes that are differentially regulated by glucocorticoids and progestins. A mouse fibroblast cell line (4F) stably expressing both GR and PR and containing a single copy of a multifunctional selection plasmid is generated. This line is transduced with a self-inactivating retroviral promoter-trap vector carrying coding sequences for Cre-recombinase (Cre) in the U3 region. Integration of the provirus places Cre expression under the control of a genomic flanking sequence. Activation of Cre expression from integration into active genes results in a permanent switch between the selectable marker genes that converts the cells from neomycin-resistant to hygromycin-resistant. Selection for hygromycin resistance after hormone treatment yields recombinants in which Cre sequences in the U3 region are expressed from hormone-inducible upstream cellular promoters. Because Cre-mediated recombination is a permanent event, the expression of the selectable marker genes is independent of ongoing Cre expression. Thus this system permits the identification of genes that are transiently or weakly induced by hormone.

Comparison of three selectable marker genes for transformation of wheat by microprojectile bombardment

1998 ◽  
Vol 25 (1) ◽  
pp. 39 ◽  
Author(s):  
Barbara Witrzens ◽  
Richard I.S. Brettell ◽  
Fiona R. Murray ◽  
David McElroy ◽  
Zhongyi Li ◽  
...  
Keyword(s):  
Selectable Marker ◽  
Microprojectile Bombardment ◽  
Single Copy ◽  
Transgenic Wheat ◽  
Immature Embryos ◽  
Marker Genes ◽  
Hygromycin B ◽  
Neomycin Phosphotransferase ◽  
Multiple Copy ◽  
Selectable Marker Genes

Three selectable marker genes were compared for their efficacy in the production of transgenic wheat plants following microprojectile bombardment of cultured immature embryos. While transformed plants were recovered using the bar (phosphinothricin acetyltransferase) gene in combination with bialaphos, and the aphA (neomycin phosphotransferase) gene in combination with geneticin or paromomycin, no transgenic material was obtained with the hpt (hygromycin phosphotransferase) gene and hygromycin B. Southern analysis revealed single copy as well as multiple copy insertions of the bar and aphA transgenes. Inheritance of these selectable marker genes was demonstrated in the T1 generation progenies.

Positive selectable marker genes for routine plant transformation

2002 ◽  
Vol 38 (2) ◽  
pp. 125-128 ◽  
Author(s):  
Suprasanna Penna ◽  
László Sági ◽  
Rony Swennen
Keyword(s):  
Plant Transformation ◽  
Selectable Marker ◽  
Marker Genes ◽  
Selectable Marker Genes

scholarly journals New chimeric promoter useful for expression of selectable marker genes in rice transformation

2005 ◽  
Vol 22 (4) ◽  
pp. 287-294
Author(s):  
Hiromi Higo ◽  
Kayo Tsuruya ◽  
Hironori Mano ◽  
Kana Hasegawa ◽  
Yuzo Minobe
Keyword(s):  
Selectable Marker ◽  
Marker Genes ◽  
Rice Transformation ◽  
Chimeric Promoter ◽  
Selectable Marker Genes

scholarly journals Partial Purification and Characterization of Biological Effects of a Lipid Toxin Produced byMycobacterium ulcerans

1998 ◽  
Vol 66 (2) ◽  
pp. 587-593 ◽  
Author(s):  
Kathleen M. George ◽  
Lucia P. Barker ◽  
Diane M. Welty ◽  
P. L. C. Small
Keyword(s):  
Culture Supernatant ◽  
Cytopathic Effect ◽  
Biological Effects ◽  
Buruli Ulcer ◽  
Fibroblast Cell ◽  
Mouse Fibroblast ◽  
Partial Purification ◽  
Mycobacterial Diseases ◽  
Mouse Fibroblast Cell Line

ABSTRACT Organisms in the genus Mycobacterium cause a variety of human diseases. One member of the genus, M. ulcerans, causes a necrotizing skin disease called Buruli ulcer. Buruli ulcer is unique among mycobacterial diseases in that the organisms at the site of infection are extracellular and there is little acute inflammatory response. Previous literature reported the presence of a toxin in the culture supernatant of M. ulcerans which causes a cytopathic effect on the mouse fibroblast cell line L929 in which the adherent cells round up and detach from the tissue culture plate. Here we report partial purification of a lipid toxin from the culture supernatant of M. ulcerans which is capable of causing the cytopathic effect on L929 cells. We also show that this cytopathic effect is a result of cytoskeletal rearrangement. The M. ulcerans toxin does not cause cell death but instead arrests cells in the G1 phase of the cell cycle.

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