10.25402/BTN.11949801.v1
Min Li
Min
Li
Ling-Yan Jiang
Ling-Yan
Jiang
Qin Liu
Qin
Liu
Yuan-Hang Wu
Yuan-Hang
Wu
Guo-Dao Liu
Guo-Dao
Liu
Yin-Hua Chen
Yin-Hua
Chen
Li-Juan Luo
Li-Juan
Luo
A Method Combining TA Cloning and Fluorescence Screening for Rapid Acquisition of Transgenic Seeds
Future Science Group
2020
Arabidopsis thaliana
Stable expression vector
OLE1 gene
TA cloning
Green fluorescent protein (GFP)
Selection marker
Plant Biology not elsewhere classified
Gene Expression (incl. Microarray and other genome-wide approaches)
2020-03-06 14:17:31
Figure
https://future-science-group.figshare.com/articles/figure/A_Method_Combining_TA_Cloning_and_Fluorescence_Screening_for_Rapid_Acquisition_of_Transgenic_Seeds/11949801
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<p><b>Fig S1 T</b><b>he picture of Gel electrophoresis during the
constraction of pOGT vector</b><b></b></p>
<p>(a) <a>Gel electrophoresis</a> of PCR amplified <i>Pro<sub>OLE1</sub>::OLE1</i>
fragment (lane 1; amplified fragment: 2245 bp, indicated by arrow). (b) <i>Xho</i>Ⅰ
and <i>Sal</i>Ⅰ digestion of the vector pA7-GFP (lane 2, 3; large fragment:
4591 bp; small fragment: 57 bp; both fragments were indicated by arrows). (c)
Colony PCR from five random clones using primers pOLE1-F and pOLE1-R (lane
4-8; amplified fragment: 2245 bp; indicated by arrow). (d) Gel electrophoresis
of PCR amplified <i>Pro<sub>OLE1</sub>::OLE1-GFP</i> fragment (lane 9,
amplified fragment: 2989 bp; indicated by arrow). (e) <i>Hin</i>dⅢ digestion
of the vector pCXSN (lane 10, 11; fragment: 10802 bp; indicated by arrow). (f)
<i>Hin</i>dIII digestion of pCXSN-OLE1-GFP (lane 12, 13; large fragment:
10802 bp; small fragment: 2957 bp; both fragments were indicated by arrows ).
(g) <i>Xcm</i>I digestion of the vector pOGT with mutated promoter and the
pCXSN-OLE1-GFP with original promoter. The asterisks indicated the resulting
fragments after <i>Xcm</i>I digestion. The last lane was the undigested pOGT
vector. </p>
<p><b>Fig S2 PCR amplification of <i>GFP</i>, <i>HygR</i>, <i>ACT</i>2
fragment from T<sub>1</sub> transgenic plants expressing pOGT. </b><b></b></p>
<p>Lane 1-20 shows the amplification from 20 independent T<sub>1</sub>
plants. Lane 21 shows the amplification from <a>Arabidopsis wild
type plants</a> (Col-0). Lane 22 shows the amplification with no template.
Lane 23 shows the amplification from pOGT plasmid. <i>GFP</i><a> fragment</a>: 272 bp, <i>HygR</i> fragment: 444 bp, <i>ACT2</i>
fragment: 188 bp.</p>
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