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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

Rhizome Project Meeting(2)5-4-2011

Min-Jeong Kim

Gang Lab

Institute of Biological Chemistry

Washington State University

Pullman, WA


Rhizome project meeting 2 5 4 2011

Why Johnsongrass?


Rhizome project meeting 2 5 4 2011

Johnsongrass

www.usda.gov


Distribution of johnsongrass

Distribution of Johnsongrass

www.usda.gov


Rhizome project meeting 2 5 4 2011

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


Rhizome project meeting 2 5 4 2011

0 week

1 week

2 weeks

4 weeks


Expression analysis of knox transcription factors based on ginger and turmeric microarray data

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


Knox 1

Knox (1)

Intensity


Knox 2

Knox (2)

Intensity


Knox 3

Knox (3)

Intensity


Knox 4

Knox (4)

Intensity


Knox 5

Knox (5)

Intensity


Alignment of knox genes in sorghum bicolor 1

Alignment of Knox genes in Sorghum bicolor (1)


Alignment of knox genes in sorghum bicolor 2

Alignment of Knox genes in Sorghum bicolor (2)


Rhizome project meeting 2 5 4 2011

Alignment of Knox genes in Sorghum bicolor (3)


Alignment of knox genes in sorghum bicolor 4

Alignment of Knox genes in Sorghum bicolor (4)


Map of pk7gwiwg2 ii rnai vector

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 on Selection Media


Transgenic plants in the greenhouse

Transgenic Plants in the Greenhouse


Insert confirmation 1

Insert Confirmation (1)

M

PC

NT

2

6

10

M

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


Insert confirmation 2

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

Thank You


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