Rhizome project meeting 2 5 4 2011
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Rhizome Project Meeting(2) 5-4-2011. Min- Jeong Kim Gang Lab Institute of Biological Chemistry Washington State University Pullman, WA. Why Johnsongrass ?. Johnsongrass. www.usda.gov. Distribution of Johnsongrass. www.usda.gov. Work Flow. Total RNA. cDNA synthesis.

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Rhizome Project Meeting(2) 5-4-2011

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Rhizome Project Meeting(2)5-4-2011

Min-Jeong Kim

Gang Lab

Institute of Biological Chemistry

Washington State University

Pullman, WA


Why Johnsongrass?


Johnsongrass

www.usda.gov


Distribution of Johnsongrass

www.usda.gov


Work Flow

Total RNA

cDNA synthesis

PCR with gene-specific primers

LR rxn with pK7GWIWG2(II)

Sequence confirmation

Cloning into pCR8/GW/TOPO vector

Sequence confirmation

Transfer into LBA4404 Agro. strain

Plant transformation


0 week

1 week

2 weeks

4 weeks


Expression Analysis ofKnox Transcription Factors(Based on Ginger and Turmeric Microarray Data)


Knox (1)

Intensity


Knox (2)

Intensity


Knox (3)

Intensity


Knox (4)

Intensity


Knox (5)

Intensity


Alignment of Knox genes in Sorghum bicolor (1)


Alignment of Knox genes in Sorghum bicolor (2)


Alignment of Knox genes in Sorghum bicolor (3)


Alignment of Knox genes in Sorghum bicolor (4)


Map of pK7GWIWG2(II) RNAi Vector

Karimi, M., Inzé, D., Depicker, A., Gateway vectors for Agrobacterium-mediated plant transformation. Trends Plant Sci. 2002 May;7(5): 193-195


Transgenic Plants on Selection Media


Transgenic Plants in the Greenhouse


Insert Confirmation (1)

M

PC

NT

2

6

10

M

M, marker; PC, positive control (vector); NT, non-transgenic


Insert Confirmation (2)

M

PC

NT

2

6

10

12

16

M

NC

M, size marker; PC, positive control (vector); NT, non-transgenic;

NC, negative control (without PCR template)


Thank You


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