Vectors

Understanding regulation of developmental events has increasingly required the use of tissue-specific expression of diverse factors affecting plant growth and environmental responses. 

To allow for cloning of presumptive promoters with tissue-specific activities, we created two plant expression vectors with multiple cloning sites upstream of a Gateway cassette for expression of either untagged or YFP-tagged genes of interest (Michniewicz et al., 2015) by modifying the pEarleyGate100 and pEarleyGate104 constructs (Earley et al., 2006).  For fast and easy tissue-specific expression of desired genes, we further developed a set of Gateway-compatible tissue-specific gene expression vectors that allow for the expression of YFP-tagged or untagged proteins driven by various promoters (see table below).  Additional vectors will be added to this table as they are created.  These vectors provide an invaluable resource to the plant community, allowing for rapid generation of a variety of tissue-specific expression constructs.

If you use these vectors in published work, please cite:
Michniewicz, M, EM Frick, and LC Strader (2015) Gateway-compatible tissue-specific vectors for plant transformation.  BMC-Research Notes, in revision.

If you have created additional expression vectors using the pMCS:GW or pMCS:YFP-GW backbone, please let us  know.  We would be happy to include these vectors in our table.

Vector Name

Expected tissue expression

Vector sequence

ABRC Stock #

Reference

pMCS:GW

none; multiple cloning site for promoter cloning

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CD3-1933

 

Michniewicz et al., 2015

pMCS:YFP-GW

none; multiple cloning site for promoter cloning

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CD3-1934 

Michniewicz et al., 2015

pADH1:GW

root, anoxic tissues (Dolferus et al., 1994; Chung and Ferl, 1999)

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CD3-1935 

Michniewicz et al., 2015

pADH1:YFP-GW

root, anoxic tissues (Dolferus et al., 1994; Chung and Ferl, 1999)

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CD3-1936 

Michniewicz et al., 2015

pCAB1:GW

shoot, photosynthetic tissues( Karlin-Neumann et al., 1988; Gao and Kaufman, 1994)

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CD3-1937 

Michniewicz et al., 2015

pCAB1:YFP-GW

shoot, photosynthetic tissues( Karlin-Neumann et al., 1988; Gao and Kaufman, 1994)

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CD3-1938 

Michniewicz et al., 2015

pCOBL1:GW

lateral root primordia, columella, leaf vascular tissue and hydathodes (Brady et al., 2007)

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CD3-1939 

Michniewicz et al., 2015

pCOBL1:YFP-GW

lateral root primordia, columella, leaf vascular tissue and hydathodes (Brady et al., 2007)

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CD3-1940 

Michniewicz et al., 2015

 

    

pEXP7:GW

root trichoblast (Cho and Cosgrove, 2002)

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 CD3-1941

Michniewicz et al., 2015

pEXP7:YFP-GW

root trichoblast (Cho and Cosgrove, 2002)

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CD3-1942 

Michniewicz et al., 2015

 

    

pLBD16:GW

lateral root primordia (Shuai et al., 2002; Laplaze et al., 2005; Laplaze et al., 2007)

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CD3-1943 

Michniewicz et al., 2015

pLBD16:YFP-GW

lateral root primordia (Shuai et al., 2002; Laplaze et al., 2005; Laplaze et al., 2007)

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CD3-1944 

Michniewicz et al., 2015

pSCR:GW

root endodermis, endodermis initials, quiescent center, shoot apical meristem L1 layer, shoot endodermis, shoot tissue surrounding vascular bundles (Di Laurenzio et al., 1996; Malamy and Benfey, 1997; Wysocka-Diller et al., 2000)

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CD3-1945 

Michniewicz et al., 2015

pSCR:YFP-GW

root endodermis, endodermis initials, quiescent center, shoot apical meristem L1 layer, shoot endodermis, shoot tissue surrounding vascular bundles (Di Laurenzio et al., 1996; Malamy and Benfey, 1997; Wysocka-Diller et al., 2000)

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CD3-1946 

Michniewicz et al., 2015

pUBQ10:GW

throughout the plant (Czechowski et al., 2005)

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CD3-1947 

Michniewicz et al., 2015

pUBQ10:YFP-GW

throughout the plant (Czechowski et al., 2005)

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CD3-1948 

Michniewicz et al., 2015

pWOL:GW

root vascular cylinder and pericycle (Mähönen et al., 2000; Bonke et al., 2003)

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CD3-1949 

Michniewicz et al., 2015

pWOL:YFP-GW

root vascular cylinder and pericycle (Mähönen et al., 2000; Bonke et al., 2003)

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CD3-1950 

Michniewicz et al., 2015

 

The pMCS:GW vector consists of a T-DNA left border, followed by the Basta herbicide resistance gene, a multiple cloning site, the Gateway cassette (attR1, chloramphenicol resistance gene, ccdB, attR2), the 3' sequence of the octopine synthase gene, and T-DNA right border.  

The pMCS:YFP-GW vector consists of a T-DNA left border, followed by the Basta herbicide resistance gene, a multiple cloning site, the in-frame coding region for yellow fluorescent protein (YFP), the Gateway cassette (attR1, chloramphenicol resistance gene, ccdB, attR2), the 3' sequence of the octopine synthase gene, and T-DNA right border for creation of N-terminal YFP fusion proteins. 

pMCS:GW, pMCS:YFP-GW, and variants thereof confer kanamycin resistance to Escherichia coli and Agrobacterium tumefaciens.  The T-DNA from these vectors confers phosphinothricin (Basta) resistance to plants.