RNA Extraction by the Guanidine Thiocyanate Procedure

McGookin, R.

doi:10.1385/0-89603-064-4:113

Cited by 32 publications

(25 citation statements)

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“…Total RNA was isolated from Arabidopsis seedlings using the guanidine thiocyanate method (McGookin, 1984). First-strand cDNA was synthesized from 2.5 mg total RNA using Moloney murine leukemia virus reverse transcriptase (ReverScript I; Nippon Gene, Tokyo) and subjected to PCR.…”

Section: Rna Isolation and Rt-pcrmentioning

confidence: 99%

Brassinosteroid Homeostasis in Arabidopsis Is Ensured by Feedback Expressions of Multiple Genes Involved in Its Metabolism

et al. 2005

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Homeostasis of brassinosteroids (BRs) is essential for normal growth and development in higher plants. We examined responsiveness of 11 BR metabolic gene expressions to the decrease or increase of endogenous BR contents in Arabidopsis (Arabidopsis thaliana) to expand our knowledge of molecular mechanisms underlying BR homeostasis. Five BR-specific biosynthesis genes (DET2, DWF4, CPD, BR6ox1, and ROT3) and two sterol biosynthesis genes (FK and DWF5) were upregulated in BR-depleted wild-type plants grown under brassinazole, a BR biosynthesis inhibitor. On the other hand, in BRexcessive wild-type plants that were fed with brassinolide, four BR-specific synthesis genes (DWF4, CPD, BR6ox1, and ROT3) and a sterol synthesis gene (DWF7) were down-regulated and a BR inactivation gene (BAS1) was up-regulated. However, their response to fluctuation of BR levels was highly reduced (DWF4) or nullified (the other eight genes) in a bri1 mutant. Taken together, our results imply that BR homeostasis is maintained through feedback expressions of multiple genes, each of which is involved not only in BR-specific biosynthesis and inactivation, but also in sterol biosynthesis. Our results also indicate that their feedback expressions are under the control of a BRI1-mediated signaling pathway. Moreover, a weak response in the mutant suggests that DWF4 alone is likely to be regulated in other way(s) in addition to BRI1 mediation.

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Section: Rna Isolation and Rt-pcrmentioning

confidence: 99%

Brassinosteroid Homeostasis in Arabidopsis Is Ensured by Feedback Expressions of Multiple Genes Involved in Its Metabolism

et al. 2005

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“…Total RNA was prepared from various maize tissues by the guanidine thiocyanate procedure (McGookin, 1984). The RNA was subjected to electrophoresis on a 1.2% (w/v) agarose gel (Sambrook et al, 1989), and blotted onto nylon membranes (Hybond-N1; Amersham Biosciences, Buckinghamshire, UK).…”

Section: Northern Analysismentioning

confidence: 99%

Molecular Characterization of Cytokinin-Responsive Histidine Kinases in Maize. Differential Ligand Preferences and Response to cis-Zeatin

et al. 2004

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Genes for cytokinin-responsive His-protein kinases (ZmHK1, ZmHK2, and ZmHK3a) were isolated from maize (Zea mays). Heterologous expression of each of the ZmHKs in Escherichia coli having the DrcsC and cps::lacZ genetic background conferred cytokinin-inducibility of lacZ expression on the bacteria. In the recombinant E. coli system, ZmHK1 and ZmHK3a were more sensitive to free-base cytokinins than to the corresponding nucleosides; isopentenyladenine was most effective for ZmHK1, while ZmHK2 tended to be most sensitive to trans-zeatin and the riboside. In contrast to a known cytokinin receptor of Arabidopsis (AHK4/CRE1/WOL), all ZmHKs responded to cis-zeatin (cZ), which generally is believed to be inactive or only weakly active. In cultured maize cells, expression of ZmRR1, a cytokinin-inducible response regulator, was induced by cZ as well as by trans-zeatin. These results strongly suggest that maize cytokinin receptors differ in ligand preference, and that cZ is an active cytokinin at least in maize.

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“…Total RNA was prepared from the 19-h-greened maize leaves by a guanidine thiocyanate procedure (McGookin 1984). Poly(A) RNA was purified with Oligotex-dT30 (Takara Bio Inc., Shiga).…”

Section: Construction Of Cdna Library and Isolation Of Cdnasmentioning

confidence: 99%

“…Maize plants were grown hydroponically in Arnon and Hoagland solution for 14 d and total RNA was isolated from their roots and leaf blades by a guanidine thiocyanate procedure (McGookin 1984). First-strand cDNA was synthesized with an oligo(dT) primer using an AMV Reverse Transcriptase First-strand cDNA Synthesis Kit (Life Sciences, Inc., St. Petersburg, FL, U.S.A.).…”

Section: Rt-pcr Analysismentioning

confidence: 99%

Differentiation of Dicarboxylate Transporters in Mesophyll and Bundle Sheath Chloroplasts of Maize

Taniguchi

Nagasaki

Kawasaki

et al. 2004

Plant and Cell Physiology

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;In NADP-malic enzyme-type C 4 plants such as maize (Zea mays L.), efficient transport of oxaloacetate and malate across the inner envelope membranes of chloroplasts is indispensable. We isolated four maize cDNAs, ZmpOMT1 and ZmpDCT1 to 3, encoding orthologs of plastidic 2-oxoglutarate/malate and general dicarboxylate transporters, respectively. Their transcript levels were upregulated by light in a cell-specific manner; ZmpOMT1 and ZmpDCT1 were expressed in the mesophyll cell (MC) and ZmpDCT2 and 3 were expressed in the bundle sheath cell (BSC). The recombinant ZmpOMT1 protein expressed in yeast could transport malate and 2-oxoglutarate but not glutamate. By contrast, the recombinant ZmpDCT1 and 2 proteins transported 2-oxoglutarate and glutamate at similar affinities in exchange for malate. The recombinant proteins could also transport oxaloacetate at the same binding sites as those for the dicarboxylates. In particular, ZmpOMT1 transported oxaloacetate at a higher efficiency than malate or 2-oxoglutarate. We also compared the activities of oxaloacetate transport between MC and BSC chloroplasts from maize leaves. The K m value for oxaloacetate in MC chloroplasts was one order of magnitude lower than that in BSC chloroplasts, and was close to that determined with the recombinant ZmpOMT1 protein. Southern analysis revealed that maize has a single OMT gene. These findings suggest that ZmpOMT1 participates in the import of oxaloacetate into MC chloroplasts in exchange for stromal malate. In BSC chloroplasts, ZmpDCT2 and/or ZmpDCT3 were expected to import malate that is transported from MC.

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RNA Extraction by the Guanidine Thiocyanate Procedure

Cited by 32 publications

References 0 publications

Brassinosteroid Homeostasis in Arabidopsis Is Ensured by Feedback Expressions of Multiple Genes Involved in Its Metabolism

Brassinosteroid Homeostasis in Arabidopsis Is Ensured by Feedback Expressions of Multiple Genes Involved in Its Metabolism

Molecular Characterization of Cytokinin-Responsive Histidine Kinases in Maize. Differential Ligand Preferences and Response to cis-Zeatin

Differentiation of Dicarboxylate Transporters in Mesophyll and Bundle Sheath Chloroplasts of Maize

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