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

Producción científica: Article › revisión exhaustiva

7 Citas (Scopus)

Resumen

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.

Idioma original	English
Páginas (desde-hasta)	425-432
Número de páginas	8
Publicación	Biochemical and Biophysical Research Communications
Volumen	160
N.º	2
DOI	https://doi.org/10.1016/0006-291X(89)92450-9
Estado	Published - abr 28 1989

Nota bibliográfica

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.

Financiación

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.

Financiadores	Número del financiador
Tobacco and Health Research Institute	4E021
U.S. Department of Agriculture	85-CRCR-1-1536

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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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.",

year = "1989",

month = apr,

day = "28",

doi = "10.1016/0006-291X(89)92450-9",

language = "English",

volume = "160",

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T1 - Expression of a potyvirus non-structural protein in transgenic tobacco

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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UR - https://www.scopus.com/pages/publications/0024980005#tab=citedBy

U2 - 10.1016/0006-291X(89)92450-9

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

M3 - Article

C2 - 2541699

AN - SCOPUS:0024980005

SN - 0006-291X

VL - 160

SP - 425

EP - 432

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

IS - 2

ER -

Expression of a potyvirus non-structural protein in transgenic tobacco

Resumen

Nota bibliográfica

Financiación

ASJC Scopus subject areas

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