Expression of a potyvirus non-structural protein in transgenic tobacco

Robert Graybosch; Gary M. Hellmann; John G. Shaw; Robert E. Rhoads; Arthur G. Hunt

doi:10.1016/0006-291X(89)92450-9

Expression of a potyvirus non-structural protein in transgenic tobacco

Robert Graybosch, Gary M. Hellmann, John G. Shaw, Robert E. Rhoads, Arthur G. Hunt

Research output: Contribution to journal › Article › peer-review

7 Scopus citations

Abstract

A cDNA fragment encoding the cytoplasmic inclusion protein of tobacco vein mottling virus was inserted into the plant expression cassette of a Ti plasmid-based binary vector. The vector was transferred to Agrobacterium tumifaciens, and following a modified leaf disc procedure, transformed tobacco plants were obtained. Analysis of poly(A)⁺ RNA from transgenic plants revealed a novel RNA of approximately 2100 nucleotides possessing tobacco vein mottling virus sequences. Also, immunoprecipitation of protein extracts of [³⁵S]methionine-labeled transformed callus using anti-cytoplasmic inclusion protein antiserum revealed a polypeptide of approximately 70 kDa. This size is consistent with that predicted from the inserted tobacco vein mottling virus coding sequences. Together these data demonstrate the expression of the cytoplasmic inclusion protein in the absence of viral infections.

Original language	English
Pages (from-to)	425-432
Number of pages	8
Journal	Biochemical and Biophysical Research Communications
Volume	160
Issue number	2
DOIs	https://doi.org/10.1016/0006-291X(89)92450-9
State	Published - Apr 28 1989

Bibliographical note

Funding Information:
Acknowledgments: We thank L. Domier and K. Franklin for gifts of TVMV clones, K. Gibb and J. Murphy for assistance with the analysis for inclusion bodies, and M. MacDonald for excellent technical assistance. This work was supported by USDA grant #85-CRCR-1-1536 and Tobacco Health Research Institute Grant 4E021.

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/0006-291X(89)92450-9

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title = "Expression of a potyvirus non-structural protein in transgenic tobacco",

abstract = "A cDNA fragment encoding the cytoplasmic inclusion protein of tobacco vein mottling virus was inserted into the plant expression cassette of a Ti plasmid-based binary vector. The vector was transferred to Agrobacterium tumifaciens, and following a modified leaf disc procedure, transformed tobacco plants were obtained. Analysis of poly(A)+ RNA from transgenic plants revealed a novel RNA of approximately 2100 nucleotides possessing tobacco vein mottling virus sequences. Also, immunoprecipitation of protein extracts of [35S]methionine-labeled transformed callus using anti-cytoplasmic inclusion protein antiserum revealed a polypeptide of approximately 70 kDa. This size is consistent with that predicted from the inserted tobacco vein mottling virus coding sequences. Together these data demonstrate the expression of the cytoplasmic inclusion protein in the absence of viral infections.",

author = "Robert Graybosch and Hellmann, {Gary M.} and Shaw, {John G.} and Rhoads, {Robert E.} and Hunt, {Arthur G.}",

note = "Funding Information: Acknowledgments: We thank L. Domier and K. Franklin for gifts of TVMV clones, K. Gibb and J. Murphy for assistance with the analysis for inclusion bodies, and M. MacDonald for excellent technical assistance. This work was supported by USDA grant #85-CRCR-1-1536 and Tobacco Health Research Institute Grant 4E021.",

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AU - Graybosch, Robert

AU - Hellmann, Gary M.

AU - Shaw, John G.

AU - Rhoads, Robert E.

AU - Hunt, Arthur G.

N1 - Funding Information: Acknowledgments: We thank L. Domier and K. Franklin for gifts of TVMV clones, K. Gibb and J. Murphy for assistance with the analysis for inclusion bodies, and M. MacDonald for excellent technical assistance. This work was supported by USDA grant #85-CRCR-1-1536 and Tobacco Health Research Institute Grant 4E021.

PY - 1989/4/28

Y1 - 1989/4/28

N2 - A cDNA fragment encoding the cytoplasmic inclusion protein of tobacco vein mottling virus was inserted into the plant expression cassette of a Ti plasmid-based binary vector. The vector was transferred to Agrobacterium tumifaciens, and following a modified leaf disc procedure, transformed tobacco plants were obtained. Analysis of poly(A)+ RNA from transgenic plants revealed a novel RNA of approximately 2100 nucleotides possessing tobacco vein mottling virus sequences. Also, immunoprecipitation of protein extracts of [35S]methionine-labeled transformed callus using anti-cytoplasmic inclusion protein antiserum revealed a polypeptide of approximately 70 kDa. This size is consistent with that predicted from the inserted tobacco vein mottling virus coding sequences. Together these data demonstrate the expression of the cytoplasmic inclusion protein in the absence of viral infections.

AB - A cDNA fragment encoding the cytoplasmic inclusion protein of tobacco vein mottling virus was inserted into the plant expression cassette of a Ti plasmid-based binary vector. The vector was transferred to Agrobacterium tumifaciens, and following a modified leaf disc procedure, transformed tobacco plants were obtained. Analysis of poly(A)+ RNA from transgenic plants revealed a novel RNA of approximately 2100 nucleotides possessing tobacco vein mottling virus sequences. Also, immunoprecipitation of protein extracts of [35S]methionine-labeled transformed callus using anti-cytoplasmic inclusion protein antiserum revealed a polypeptide of approximately 70 kDa. This size is consistent with that predicted from the inserted tobacco vein mottling virus coding sequences. Together these data demonstrate the expression of the cytoplasmic inclusion protein in the absence of viral infections.

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Expression of a potyvirus non-structural protein in transgenic tobacco

Abstract

Bibliographical note

ASJC Scopus subject areas

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