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.

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.

Heat shock induced excision of selectable marker genes in transgenic banana by the Cre-lox site-specific recombination system

2012 ◽  
Vol 159 (4) ◽  
pp. 265-273 ◽  
Author(s):  
Borys Chong-Pérez ◽  
Rafael G. Kosky ◽  
Maritza Reyes ◽  
Luis Rojas ◽  
Bárbara Ocaña ◽  
...  
Keyword(s):  
Heat Shock ◽  
Selectable Marker ◽  
Marker Genes ◽  
Site Specific ◽  
Site Specific Recombination ◽  
Selectable Marker Genes ◽  
Recombination System ◽  
Transgenic Banana ◽  
Site Specific Recombination System

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 Erratum: Excision of selectable marker genes from transgenic plants

2002 ◽  
Vol 20 (8) ◽  
pp. 843-843 ◽  
Author(s):  
Peter D. Hare ◽  
Nam-Hai Chua
Keyword(s):  
Transgenic Plants ◽  
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

Generation of selectable marker-free transgenic rice using double right-border (DRB) binary vectors

2001 ◽  
Vol 28 (3) ◽  
pp. 241 ◽  
Author(s):  
Hui-Juan Lu ◽  
Xue-Rong Zhou ◽  
Zhu-Xun Gong ◽  
Narayana M. Upadhyaya
Keyword(s):  
Selectable Marker ◽  
Binary Vector ◽  
Marker Gene ◽  
Marker Genes ◽  
Binary Vectors ◽  
Selectable Marker Genes ◽  
Right Border ◽  
Marker Free ◽  
Dna Vectors ◽  
Transgenic Progeny

Currently employed transformation systems require selectable marker genes encoding antibiotic or herbicide resistance, along with the gene of interest (GOI), to select transformed cells from among a large population of untransformed cells. The continued presence of these selectable markers, especially in food crops such as rice (Oryza sativa L.), is of increasing public concern. Techniques based on DNA recombination and Agrobacterium-mediated co-transformation with two binary vectors in a single or two different Agrobacterium strains, or with super-binary vectors carrying two sets of T-DNA border sequences (twin T-DNA vectors), have been employed by researchers to produce selectable marker-free (SMF) transgenic progeny. We have developed a double right-border (DRB) binary vector carrying two copies of T-DNA right-border (RB) sequences flanking a selectable marker gene, followed by a GOI and one copy of the left border sequence. Two types of T-DNA inserts, one initiated from the first RB containing both the selectable gene and the GOI, and the other from the second RB containing only the GOI, were expected to be produced and integrated into the genome. In the subsequent generation, these inserts could segregate away from each other, allowing the selection of the progeny with only the GOI. We tested this vector using two selectable marker genes and successfully obtained progeny plants in which the second selectable marker gene segregated away from the first. Using the DRB binary vector system, we recovered SMF transgenic lines containing a rice ragged stunt virus (RRSV)-derived synthetic resistance gene in the rice cultivars Jarrah and Xiu Shui. Approximately 36–64% of the primary transformants of these cultivars yielded SMF progeny. Among SMF Jarrah transgenic progeny <50% of plants contained the RRSV transgene. Thus, we have developed an efficient vector for producing SMF plants that allows straightforward cloning of any GOIs in comparison with the published ‘twin T-DNA’ vectors.

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