The Incorporation of Amino Acids into Protein by Cell-free Extracts from Tobacco Leaves<sup>*</sup>

Spencer, Donald; Wildman, S. G.

doi:10.1021/bi00895a019

Cited by 153 publications

(57 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Disposable plastic gloves were used during chopping, and in expressing liquid from the glass wool pad to minimize contaminationl with exogenous ribonuclease (15). The homogenate was sequentially centrifuged at 1000 X g X 10 minutes, 12,000 Xg X 30 minutes, and 144,000 X g X 120 minutes (30), and pellets were resuspended in fresh medium, the same as used to prepare the mince. The resuspended 1000 X g pellet is the chloroplast preparatioli used standardly.…”

Section: Methodsmentioning

confidence: 99%

“…Assay was carried out by the procedure of Spencer and Wildman (30). Reagents other than L-leucinel4C, uniformlv labeled (New England Nuclear Corporation, Boston, Massachusetts), P-enolpyruvate (Sigma Chemical Corporation, St. Louis, Missouri), and pyruvate kinase stored frozen in snmall batches, thawed once, and the unused portion discarded.…”

Section: Methodsmentioning

confidence: 99%

“…Other reaction mixture components were the same as described under (27 Non-ionic detergents have been reported to have no effect onl bacterial growth (23). The non-ionic detergent, Triton X-100 has been reported to leave nuclei (30) and bacteria (4) unaffected, wlhile framenting chloroplasts. The usefulness of Triton X-100 to separate chloroplastic material fromn bacteria, leaf cells and nuclei of the chioroplast fraction used for these amlino acid incorporation studies was examined.…”

Section: Methodsmentioning

confidence: 99%

“…Here, chloroplast suspension was treated, or not treated with Tritoni, after 20 minutes, centrifuged at 6000 X g, and the pellets suspended, fixed, and stained in small volumes of acetocarmine. (30), or hypotonic (0.01 M tris, pH 7.8; 5n mm magnesium acetate; 3 mm mercaptoethanol) medituml was incubated with radioactive amino acid for 60 minutes. The numbers in parenthesis are the counts incorporated/assay in the absence of ribonuclease.…”

Section: Methodsmentioning

confidence: 99%

“…Chloroplast containinig fractions from leaves of higher plants (2,4,12,14,25,28,30,31 ), and from algal cells (10. 13) have been reported to incorporate amino acid inlto protein.…”

mentioning

confidence: 99%

See 4 more Smart Citations

In Vitro Protein Synthesis by Plastids of Phaseolus vulgaris. I. Localization of Activity in the Chloroplasts of a Chloroplast Containing Fraction from Developing Leaves

Parenti¹,

Margulies²

1967

Plant Physiol.

View full text Add to dashboard Cite

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…Chloroplast containinig fractions from leaves of higher plants (2,4,12,14,25,28,30,31 ), and from algal cells (10. 13) have been reported to incorporate amino acid inlto protein.…”

mentioning

confidence: 99%

See 3 more Smart Citations

In Vitro Protein Synthesis by Plastids of Phaseolus vulgaris. I. Localization of Activity in the Chloroplasts of a Chloroplast Containing Fraction from Developing Leaves

Parenti¹,

Margulies²

1967

Plant Physiol.

View full text Add to dashboard Cite

The Photochemical Systems of Photosynthesis

Nk¹

1968

Advances in Enzymology - And Related Areas of Molecular Biology

View full text Add to dashboard Cite

Isolationsbedingungen für Zellkerne aus Hefeprotoplasten

May¹,

Gottstein²

1971

J of Basic Microbiology

View full text Add to dashboard Cite

The structure of isolated nuclei from Saccharomyces fragilis protoplasts depends on the composition of the medium used for isolation. Buffered solutions of polymers as for instance Dextran, Ficoll and Polyvinylpyrrolidon a t p H 5.5-6.5 are suitable both for carefully lysing the protoplasts and for the isolation of the nuclei. The structure of isolated nuclei does not differ markedly from nuclei in living cells. Isolated nuclei do not, behave like osmometers but are swelling in buffered solution of sugars such as sucrose, mannitol and sorbitol. uber die Isolation von Zellkernen aus Hefeprotoplasten ist bisher nur von ROZIJN u. TONINO (1964) berichtet worden. Diese Zellkerne von Saecharomyces earlsbergensis wurden auf Grund von licht-und elektronenmikroskopischen Untersuchungen als morphologisch intakt bezeichnet und biochemisch bearbeitet (TONINO u. ROZIJN 1966, SILLEVIS SMITT etal. 1970, MOLENAAR et al. 1970).Das in der vorliegenden Arbeit beschriebene Isolationsverfahren weicht von der Methode ah, die von ROZIJN u. TONINO (1964) vetwendet wurde. Es ermoglicht aber die Isolation von Zellkernen, die sich morphologisch nur wenig von denen in intakten Zellen unterscheiden, wie durch licht-und elektronenmikroskopische Untersuchungen festgestellt werden konnte. Material und NethodenObjekt und Kultivierung : Die Versuche wurden mit Saccharomyces fragilis H, durchgefiihrt. Die Hefezellen einer 48stundigen Bierwurze-Schragagarkultur wurden mit einem Nahrniedium nach OHNISHI et al. (1966) abgeschwemmt, in einen 500-ml-Rundstehkolben mit 150 ml des gleichen Nahrmediums ubertragen und 7-8 Std. bei 28 "C auf einem Schutteltisch M 56, K 15, 240 U/min (VEB FANAL, Bad Frankenhausen) inkubiert. J e 0.5 ml dieser Kultnr dienten zum Beimpfen weiterer Rundstehkolben. Nach 15-17stundigem Schiitteln wurden die Hefezellen, die sich dann in der exponentiellen Wachstumsphase befanden, durch Zentrifugation (2000 g, 2 min) abgetrennt und 2mal mit dest. H,O gewaschen. Priiparation der Protoplasten: Folgendes Praparationsverfahren fand in Anlehnung an Untersuchungen von ROST u. VENNER (1965). SUOMALAINEN et al. (1966) und SCHWENCRE et al. (1969) Verwendung: 3 g gewaschene Hefezellen (Feuchtgewicht) wurden in 10 ml TRIS-EDTA-Mediuml) suspendiert, 10 niin bei 30 "C geschiittelt (Reziprokschiittler, Schuttelfrequenz 130/min) und danach 2mal mit dest. H,O gewaschen. Die Hefezellen wurden anschliel3end in 40 ml Protoplasten-Praparationsmedium (PPM)3) resuspendiert und 1) TRIS-EDTA-Medium: 100 mM TRIS, 100 mM EDTA-Na,, mit KOH auf p H 8,O ein-2) PPM: Phosphat-Citratpuffer nach MCILVAINE, p H 6,O; 0,6 M (NH,),SO,; 0,02 M 2-Mer-gestellt.captoathanol.

show abstract

The Incorporation of Amino Acids into Protein by Cell-free Extracts from Tobacco Leaves^*

Abstract: distribution of amino acid-incorporating activity in such extracts, and some of the characteristics of an active system associated with the 1000-g fraction.

Cited by 153 publications

References 10 publications

In Vitro Protein Synthesis by Plastids of Phaseolus vulgaris. I. Localization of Activity in the Chloroplasts of a Chloroplast Containing Fraction from Developing Leaves

In Vitro Protein Synthesis by Plastids of Phaseolus vulgaris. I. Localization of Activity in the Chloroplasts of a Chloroplast Containing Fraction from Developing Leaves

The Photochemical Systems of Photosynthesis

Isolationsbedingungen für Zellkerne aus Hefeprotoplasten

Contact Info

Product

Resources

About

The Incorporation of Amino Acids into Protein by Cell-free Extracts from Tobacco Leaves*

Abstract: distribution of amino acid-incorporating activity in such extracts, and some of the characteristics of an active system associated with the 1000-g fraction.

Cited by 153 publications

References 10 publications

In Vitro Protein Synthesis by Plastids of Phaseolus vulgaris. I. Localization of Activity in the Chloroplasts of a Chloroplast Containing Fraction from Developing Leaves

In Vitro Protein Synthesis by Plastids of Phaseolus vulgaris. I. Localization of Activity in the Chloroplasts of a Chloroplast Containing Fraction from Developing Leaves

The Photochemical Systems of Photosynthesis

Isolationsbedingungen für Zellkerne aus Hefeprotoplasten

Contact Info

Product

Resources

About

The Incorporation of Amino Acids into Protein by Cell-free Extracts from Tobacco Leaves^*