scholarly journals Keratin Biosynthesis II. Extraction and Characterization of Nucleic Acids from Wool Roots

1970 ◽  
Vol 23 (1) ◽  
pp. 139 ◽  
Author(s):  
BR Wilkinson
Keyword(s):  
Nucleic Acids ◽  
Ribosomal Rna ◽  
Good Yield ◽  
Two Stage ◽  
Extraction Procedures ◽  
Yield And Purity

Ribosomal RNA (rRNA), soluble RNA (sRNA), and DNA have been extracted from wool roots in good yield and purity. rRNA and sRNA were prepared by extracting wool roots with a hydrophilic salt and a phenol-cresol mixture and rRNA was selectively precipitated with m�cresol. DNA was extracted with a salt having lipophilic and chelating properties, together with the phenol-cresol mixture. Two�stage extraction procedures were essential for the preparation of stable, undegraded nucleic acids.

Characterization of Rapidly Labelled Nucleic Acids in Tissues of Plant Seedlings

1966 ◽  
Vol 21 (10) ◽  
pp. 983-992 ◽  
Author(s):  
Vera Hemleben-Vielhaben
Keyword(s):  
Nucleic Acids ◽  
Ribosomal Rna ◽  
Messenger Rna ◽  
Specific Activity ◽  
Supernatant Fraction ◽  
Nucleic Acid Metabolism ◽  
Tissue Homogenates ◽  
Fraction I ◽  
Plant Seedlings

The nucleic acid metabolism of plant tissues was examined by incubating seedlings of Phaseolus, Vicia, Pisum, and Soja or sections of them with 32P for short periods of time. The nucleic acids extracted from this material were fractionated on columns of methylated albumin coated on kieselgur, and the radioactivity and composition of the specific fractions were determined. Application of 32P to intact seedlings or to excised parts of seedlings resulted in the same pattern of labelled nucleic acids in all tissues, but the total amount of incorporated radioactivity was different. In all tissues investigated five rapidly labelled RNA fractions were found associated with the following components: (I) soluble RNAs, (II) DNA, (III—V) ribosomal RNA. Fraction I contained equally high amounts of CMP and GMP thus differing significantly from the soluble RNAs. The composition of fraction (II) which was probably bound to DNA in the form of a complex, was similar to that of fraction I provided the labelling period was short. Fractions III—V were of the ribosomal type. The rapidly labelled DNA fraction had a high guanine-cytosine content (60%) as compared with the bulk DNA (40%). Fractional centrifugation of the tissue homogenates revealed that the labelled RNA of the ribosomal type was partly associated with the ribosomes, partly with larger particles which were sedimented by low speed centrifugation. The RNA associated with the latter had a higher specific activity than the ribosomal RNA in the supernatant. Fraction I and the second component of the soluble RNA (s-2) were also confined to the sediment of larger particles. Actinomycin D (10 µg/ml) inhibited the incorporation of 32P in the nucleic acids. In chase experiments it caused a decrease in the specific activity of all RNA fractions, most prominently, however, in fraction I and II indicating their instability and resemblance to messenger RNA.

scholarly journals Characterization of cytoplasmic and nuclear genomes in the colorless alga Polytoma. II. General characterization of organelle nucleic acids.

1976 ◽  
Vol 69 (2) ◽  
pp. 371-382 ◽  
Author(s):  
C Siu ◽  
H Swift ◽  
K Chiang
Keyword(s):  
Nucleic Acids ◽  
Chlamydomonas Reinhardtii ◽  
Ribosomal Rna ◽  
Base Composition ◽  
Buoyant Density ◽  
Main Band ◽  
Major Species ◽  
A Minor ◽  
Cytoplasmic Ribosomes

Polytoma obtusum has a main band DNA (alpha) with a buoyant density in CsC1 of rho = 1.711 g/ml and a light DNA satellite (beta) with rho = 1.682 g/ml. beta-DNA was substantially enriched in a fraction containing small leucoplast fragments and some mitochondria, which was obtained in a pellet sedimenting between 3,000 g and 5,000 g. A crude mitochondrial pellet was also obtained by sedimenting at 12,000 g to recover particulates remaining in the supernate after 10 min at 5,000 g. This fraction contained a third DNA component (gamma) with rho = 1.714 g/ml. We have concluded that the leucoplasts of P. obtusum contain the beta-DNA (1.6882) and the mitochondria possess the gamma-component (1.714). Two distinct classess of ribosomes were isolated and separated by sucrose density gradients, a major 79S species and a minor species at 75S. The major species possessed the 25S and 18S ribosomal RNA (rRNA), characteristic of cytoplasmic ribosomes, and these particles co-sedimented in sucrose gradients with the 79S cytoplasmic ribosomes of Chlamydomonas reinhardtii. The minor species was present in about 2% of the total ribosomal population but showed an eight-to-ninefold enrichment in the leucoplast pellet, suggesting that it was of organelle origin. These 73S particles had RNA components migrating very closely with the 18S and 25S species of the 79S ribosomes, but the base composition of the rRNA from these two classes of ribosomes was significantly different; the rRNA from the 79S ribosomes had a G+C mole ratio of 50.0%, while the rRNA from the 73S class had a ratio of 47.5%. By comparison, chloroplast ribosomes of C. reinhardtii were found to sediment at 70S and contain rRNA molecules of 23S and 16S, with a G + C content of 51.0%. These findings support the concept that the Polytoma leucoplast possesses characteristic genetic and protein-forming systems.

Characterization of Glycosphingolipids and Their Diverse Lipid Forms through Two-Stage Matching of LC-MS/MS Spectra

2021 ◽  
Vol 93 (6) ◽  
pp. 3154-3162
Author(s):  
Laura S. Bailey ◽  
Fanran Huang ◽  
Tianqi Gao ◽  
Jinying Zhao ◽  
Kari B. Basso ◽  
...  
Keyword(s):  
Two Stage ◽  
Stage Matching

Cloning and characterization of the ribosomal RNA gene repeat from Ostertagia ostertagi

1991 ◽  
Vol 45 (2) ◽  
pp. 275-280 ◽  
Author(s):  
John B. Dame ◽  
Charles A. Yowell ◽  
Charles H. Courtney ◽  
W.Gary Lindgren
Keyword(s):  
Ribosomal Rna ◽  
Ostertagia Ostertagi ◽  
Ribosomal Rna Gene

scholarly journals Detection, Identification, and Molecular Characterization of the 16SrII-D Phytoplasmas Infecting Vegetable and Field Crops in Oman

Plant Disease ◽  
2018 ◽  
Vol 102 (3) ◽  
pp. 576-588 ◽  
Author(s):  
Ali M. Al-Subhi ◽  
Saskia A. Hogenhout ◽  
Rashid A. Al-Yahyai ◽  
Abdullah M. Al-Sadi
Keyword(s):  
Molecular Characterization ◽  
Ribosomal Rna ◽  
Faba Bean ◽  
Sequence Variation ◽  
Field Pea ◽  
16S Ribosomal Rna ◽  
Diverse Range ◽  
Field Crops ◽  
Phytoplasma Infection

Typical symptoms of phytoplasma infection were observed on 11 important crops in Oman that included alfalfa, sesame, chickpea, eggplant, tomato, spinach, rocket, carrot, squash, field pea, and faba bean. To identify the phytoplasmas in these crops, samples from infected and asymptomatic plants were collected, followed by amplifying and sequencing of the 16S ribosomal RNA, secA, tuf, imp, and SAP11 genes. We found that these sequences share >99% similarity with the peanut witches’ broom subgroup (16SrII-D). Whereas some sequence variation was found in the five genes among 11 phytoplasma isolates of different crops, all sequences grouped into one clade along with those of other phytoplasmas belonging to the 16SrII-D group. Thus, 16SrII-D phytoplasmas infect a diverse range of crops in Oman. Phytoplasmas in this group have not been reported to occur in carrot, spinach, rocket, and field pea previously. Within Oman, this is the first report of the presence of 16SrII-D phytoplasmas in tomato, spinach, rocket, carrot, squash, field pea, and faba bean. Sequences of the five genes enabled for better distinction of the 16SrII-D phytoplasmas that occur in Oman.

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