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Characterization of a maize endosperm culture expressing zein genes and its use in transient transformation assays

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Summary

An endosperm derived tissue culture of maize (Zea mays L.) variety A636 has been characterised for its ability to synthesize zein protein and respond to a zein gene regulatory element. Western analysis with zein specific antibodies revealed the distinct presence of zein proteins of the 15, 19 and 21 kDa classes in this tissue, in contrast to an embryo-derived Black Mexican Sweet variety tissue culture which exhibited no zein proteins. Transient transformation studies with a cauliflower mosaic virus 35S promoter and luciferase reporter gene construct demonstrate that protoplasts from this tissue culture, but not from the embryo-derived culture, respond positively to the potential enhancer which is located approximately 300 base pairs upstream of the coding region in most zein genes of maize, thus establishing the usefulness of this culture for studies of tissue specific gene regulation.

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Abbreviations

MS:

Murashige and Skoog

CaMV:

cauliflower mosaic virus

BMS:

Black Mexican Sweet

2,4-D:

2,4-dichlorophenoxy — acetic acid

DAP:

days after pollination

SDS:

sodium dodecyl sulfate

PAGE:

polyacrylamide gel electrophoresis

MES:

2-morpholinoethane-sulfonic acid

Hepes:

N-2-hydroxyethylpiper-azine-N′-2-ethansulfonic acid

Tris:

Tris-(hydroxymethyl)-aminomethane

EDTA:

ethyl-enediaminetetra-acetic acid disodium salt

PEG:

polyethylene glycol

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Communicated by H. Lörz

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Quayle, T.J.A., Hetz, W. & Feix, G. Characterization of a maize endosperm culture expressing zein genes and its use in transient transformation assays. Plant Cell Reports 9, 544–548 (1991). https://doi.org/10.1007/BF00232328

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  • DOI: https://doi.org/10.1007/BF00232328

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