DOI: 10.1111/j.1432-1033.1979.tb12791.x. Cell‐Free Transcription and Translation of Total Spinach Chloroplast DNA

Cell‐Free Transcription and Translation of Total Spinach Chloroplast DNA

10.1111/j.1432-1033.1979.tb12791.x

Crossref journal-article

Wiley

European Journal of Biochemistry (311)

Abstract

A highly efficient cell‐free Escherichia coli system for the coupled transcription/translation of chloroplast DNA is described. Evidence for the synthesis of more than 20 discrete size classes of [35S]‐methionine‐labelled polypeptides has been obtained by dodecylsulphate/polyacrylamide gel electrophoresis of the products formed in a reaction programmed with spinach chloroplast DNA. One of the major radioactive products co‐electrophoresed with the large subunit (Mr 55000) of ribulosebisphosphate carboxylase (fraction 1 protein). Its identity as the large subunit was established by comparing its partial proteolytic digestion products with those of purified large subunit protein. Because the level of radioactivity in many of the other polypeptides synthesized in vitro is so high, their identification likewise should be possible by comparing them with known chloroplast proteins.When spinach chloroplast DNA that had been digested with certain restriction endonucleases was used to programme the transcription/translation system, some of the polypeptides, which were observed using undigested DNA, were not detectable among the products. These results are discussed with regard to the distribution of restriction enzyme sites in the operons for some of the polypeptides, and with regard to the information they can provide on the location of genes on a restriction enzyme cleavage map of chloroplast DNA.

Bibliography

BOTTOMLEY, W., & WHITFELD, P. R. (1979). CellâFree Transcription and Translation of Total Spinach Chloroplast DNA. European Journal of Biochemistry, 93(1), 31â39. Portico.

Authors 2

Warwick BOTTOMLEY (first)
Paul R. WHITFELD (additional)

References 16 Referenced 78

10.1126/science.1108201
10.1007/BF00332900
{'key': 'e_1_2_3_4_2', 'first-page': '375', 'volume-title': 'The Lac Operon', 'author': 'Zubay G.', 'year': '1970'} / The Lac Operon by Zubay G. (1970)
10.1111/j.1432-1033.1976.tb10656.x
10.1016/0076-6879(67)12129-0
10.1016/0022-2836(73)90198-8
10.1093/nar/5.6.1741
10.1016/0003-9861(74)90012-5
10.1016/0003-2697(76)90202-5
10.1016/0003-2697(75)90521-7
10.1111/j.1432-1033.1975.tb02238.x
10.1016/0022-2836(75)90372-1
{'key': 'e_1_2_3_14_2', 'first-page': '413', 'volume-title': 'Acides Nucléiques et Synthèse des Protéines chez les Végétaux', 'author': 'Bottomley W.', 'year': '1977'} / Acides Nucléiques et Synthèse des Protéines chez les Végétaux by Bottomley W. (1977)
10.1016/0005-2787(73)90013-0
10.1016/0014-5793(76)80025-7
10.1073/pnas.74.12.5487

Dates

Type	When
Created	20 years, 5 months ago (March 3, 2005, 2:43 p.m.)
Deposited	1 year, 9 months ago (Nov. 23, 2023, 12:05 a.m.)
Indexed	1 year, 9 months ago (Nov. 23, 2023, 2:41 p.m.)
Issued	46 years, 8 months ago (Jan. 1, 1979)
Published	46 years, 8 months ago (Jan. 1, 1979)
Published Online	17 years, 2 months ago (June 28, 2008)
Published Print	46 years, 8 months ago (Jan. 1, 1979)

Funders 0

None

BibTeX

@article{BOTTOMLEY_1979, title={Cell‐Free Transcription and Translation of Total Spinach Chloroplast DNA}, volume={93}, ISSN={1432-1033}, url={http://dx.doi.org/10.1111/j.1432-1033.1979.tb12791.x}, DOI={10.1111/j.1432-1033.1979.tb12791.x}, number={1}, journal={European Journal of Biochemistry}, publisher={Wiley}, author={BOTTOMLEY, Warwick and WHITFELD, Paul R.}, year={1979}, month=jan, pages={31–39} }

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