Prof. Dr. Peterh nsel Dr. Reinard 2 Hannove ve r Bo Boston 3 - PowerPoint PPT Presentation

Prof. Dr. Peterh ä nsel Dr. Reinard 2

Hannove ve r Bo Boston 3

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- global issue - harmful to animals and human - dose response effect - accumulation 5

- cleaning nature by using nature itself - equip plants with a protein which binds - four different heavy metals - significant reduction 6

signal sequence (Exp4) Cellulose Binding Domain T op 4 M etal B inding P rotein 7

Plant t Vector tor 8

- a new plant vector for iGEM 9 (plasmid map originally taken from www.snapgene.com/resources)

promoter exchange: enTCUP2 promoter into 2x35s promoter removal of restriction sites: XhoI and BglII 10

GFP 11

evaluate: - the signal sequence (Exp4) - cellulose-binding domain (CBD) 12

Exponential Megaprimer PCR R1 R2 EXP CBD Term 13

pORE_E3_2 _E3_2x35 x35S_G S_GFP FP 14

transient transformation expression of p rotein signal sequence 15

Protein ein 16

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Nicotiana tabacum 18

- time - tag Nicotiana tabacum - amount of protein E. coli 19

E. coli Anti-6xHistidine Tag Antibody 20

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seeds were harvested 22

selective medium 23

heterologous expression in E. coli binding of Zn, Cd and Cu regeneration of potentially transgenic plants 24

Teabag 0 25

Hannove ve r Bo Boston 26

Exponential Megaprimer PCR 1 st step 2 nd step Ulrich et al. PLoS One. 2012;7(12)