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We modified a binary plasmid containing the complete CMV RNA 3 cDNA to facilitate insertion of desired sequences, and to give modifications of the subgenomic mRNA 4 leader sequence yielding several variants.
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We thank F. Brandizzi and C. Hawes (Oxford Brookes University) for making available the binary plasmid containing ST-mRFP fusion.
The transgenic tomato plants used in this work were generated via Agrobacterium tumefaciens-mediated transformation [ 10] with a binary plasmid containing in its T-DNA an expression cassette for the TSWV-N gene, together with a second cassette for expression of the nptII gene, as selectable marker [ 11].
Moreover, Ding and coworkers designed a binary cotransfection assay system consisting of both a donor plasmid containing the transposon and a helper plasmid containing the transposase.
Construction of a Plasmid Containing the Gene Encoding His6-TaGene Encoding
Using a yeast plasmid containing the entire coding region of STE5.
A plasmid containing the CCAAT/enhancer-binding protein (C/EBP alpha) gene transcriptionally controlled by the metallothionein promoter was constructed.
This plasmid contains the HIS3 marker gene.
Standard curves were obtained using serial dilutions of linearized plasmids containing the cloned genes (efficiency: 89 99%).
Plasmids containing the complete coding sequences of Masp1 (BC131638), Masp2 (BC013893.1) and C3 (BC043338.1) were purchased from OpenBiosystems (ThermoScientific) and subcloned into pCAGGS plasmid.
Plasmids containing the correct mutations were further verified by sequencing.
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