Sentence examples for binary vectors using the from inspiring English sources
Arabidopsis plants were transformed as described previously [ 19, 56] with Agrobacterium tumefaciens strains GV3101 pMP90 [ 101] or LBA4404 [ 102] containing the GFP binary vectors using the standard floral dip method [ 103].
Cloned genes were subcloned in the pCAMBIA1300 binary vector using the ApaI restriction site and used for Agrobacterium-mediated transformation.
Among the factors involved included the genotype of plants, types and age of tissues used, the Agrobacterium strains and binary vectors used, and the various tissue culture conditions [ 6].
The binary vectors used all contained the NPTII gene as selectable marker but differed in backbone (origin) and reporter genes or genes-of-interest involved in the regulation of seed oil composition.
Next, we constructed a set of binary transformation vectors using the transient expression vectors as backbones.
A summary of the selection process is shown in Fig. 1a, and the binary vectors used to create these transgenic lines are shown in Fig. 1b.
Transferred DNA was UV crosslinked and subsequently immobilized by baking at 120 °C for 30 min. Restricted and immobilized genomic DNA was then allowed to hybridize with DIG-labelled probes targeted against a hygromycin phosphotransferase coding sequence present towards the left border in T-DNA of the three binary vectors used to transform banana cells.
(a) Schematic representation of T-DNA regions of the CRISPR/Cas9 binary vectors used in this study.
Binary vectors used to transform Arabidopsis have been described previously [8].
The generated 126bp-fragments were cloned into the binary vector using HindIII and PstI restriction sites, 5' of the minimal promoter in pGreen- m35S::GFP.
The DNA sequence encoding 19E5scFv-GFP11 wamplifiedied by PCR and subcloned into the pENTR/D-TOPO vector (Thermo Fisher Scientific, http://www.thermofisher.co.jp/), and then recombined into the pMDC32 binary vector using LR Clonase II (Thermo Fisher Scientific).
