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Sequence Results. T he A rabidopsis I nformation R esource. The National Center for Genome Resources (NCGR), Santa Fe, New Mexico The Arabidopsis Biological Resource Center (ABRC), Ohio State University Carnegie Institution, Department of Plant Biology, Stanford University .

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The Arabidopsis Information Resource

The National Center for Genome Resources (NCGR), Santa Fe, New Mexico

The Arabidopsis Biological Resource Center (ABRC), Ohio State University

Carnegie Institution, Department of Plant Biology, Stanford University

Mission Summary: Provide research resources for the

Arabidopsis research community

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NSF 2010 Project

To determine the function of all genes in

Arabidopsis thaliana by the year 2010

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How To Determine Gene Function?

• Gene Overview

• Knockouts

• Localization

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P450s are membrane-associated proteins, either in the inner membrane of

mitochondria or in the endoplasmic reticulum of cells where they metabolize

thousands of endogenous and exogenous compounds.

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What does the number At1g11680 mean?

At (Arabidopsis thaliana)

1,2,3,4,5 (chromosome number) or M for mitochondrial or C for chloroplast.

g (gene), other letters possible for repeats etc.)

12300 (five-digit code, numbered from top/north to bottom/south of chromosome)

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Cre-loxP Site-Specific Recombination

(cyclization recombination)

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Cre recombinase (Cre):

A type I topoisomerase from P1 bacteriophage that catalyzes site-specific

recombination of DNA between loxP sites

The loxP recognition element:

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pUNI pHOST 10x BufferWaterCre

1µl1µl2µl15µl1µl

37°C

Transform in DH5α.Select on Amp.

20 min

20µl

Qiagen miniprep DNA, approx. 200 ng/µl

Recommended reaction for Cre-LoxP

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β-Glucuronidase

Green Fluorescent Protein

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β-Glucuronidase

General localization.

X-Gluc + β-Glucuronidase =

Blue

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Green Fluorescent Protein

General localization and

subcellular localization.

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Luciferase

Evaluation of gene expression/induction.

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In Summary:

The Arabidopsis Information Resource

• The primary goal of TAIR is to assist the Arabidopsis community

discover gene function.

• Extensive gene knockout and localization tools (T-DNA lines, full-length cDNAs,

and pUNI cDNAs) serve as the core elements of what TAIR offers.

• The TAIR website, its databases, and resources continue to expand, so

if you are unable to find a T-DNA insertion today…try tomorrow!

Syngenta Inc has now donated about 9,000 more lines to ABRC and

sequences to this site at Mar. 31 2006

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Key References

Cre-loxP recombination vectors for the expression of Riken Arabidopsis full-length cDNAs in plants Toshiro Shigaki, Mamata Kole, John M. Ward, Heven Sze, and Kendal D. Hirschi

BioTechniques Vol. 39, No. 3: pp 301-303 (Sep 2005)

The Cre-loxP recombination-based reporter system for plant transcriptional expression studies

Shigaki, T., R. Ravindranadha, A.B. Sivitz, J.M. Ward, H. Sze, and K.D. Hirschi

Plant Mol. Biol. 58:65-73 (2005)

The univector plasmid-fusion system, a method for rapid construction of recombinant DNA without

restriction enzymes

Liu, Q., M.Z. Li, D. Leibham, D. Cortez, and S.J. Elledge

Curr. Biol. 8:1300-1309 (1998)

A structural view of Cre-loxP site-specific recombination

Gregory D. Van Duyne

Annual Review of Biophysics and Biomolecular Structure Vol. 30: 87-104 (2001)