Isolation, characterization and expression of a cDNA encoding an elongation factor-1alpha from Porphyra yezoensis (Bangiales, Rhodophyta)

Fukuda, Satoru; Ootsuka, Shuuji; Kitade, Yukihiro; Watanabe, Toshiki; Saga, Naotsune

doi:10.1111/j.1440-1835.2002.tb00131.x

Cited by 5 publications

(5 citation statements)

References 19 publications

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“…The analysis of target gene expression at different life-history stages is an increasingly significant field of life science research [11], [18], [19]. In order to conduct a gene expression assay it is crucial to choose a suitable internal control gene that is expressed stably and, thus, the selection of this internal control gene is an absolute prerequisite for accurate quantification of target gene expression [20]–[22].…”

Section: Introductionmentioning

confidence: 99%

Variation of Expression Levels of Seven Housekeeping Genes at Different Life-History Stages in Porphyra yezoensis

Huang

et al. 2013

PLoS ONE

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In order to identify the optimal internal control for relative real-time PCR when studying target gene expression in the red alga Porphyra yezoensis, we quantified the expression of seven housekeeping genes (18S ribosomal RNA, 30S ribosomal protein S8, Polyubiquitin-2, Glyceraldehyde-3-phosphate dehydrogenase, Elongation factor 1-alpha, Beta-tubulin and Actin 3) at different life-history stages. Absolute quantification was done by normalization to total RNA quantity and by normalization to genomic DNA quantity. We used these two normalization approaches, comparing the differences of expression levels of all candidate housekeeping genes between any two generations and across three life-history stages (filamentous sporophytes, leafy gametophytes and conchospores). We found GAPDH had the best stability in all cases and we recommend that GAPDH be considered as a potential internal control for gene expression studies at different life-history stages in P. yezoensis.

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Section: Introductionmentioning

confidence: 99%

Variation of Expression Levels of Seven Housekeeping Genes at Different Life-History Stages in Porphyra yezoensis

Huang

et al. 2013

PLoS ONE

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“…The putative polyadenylation signal, CAYTG, is often found in P. yezoensis cDNA clones reported so far 3,15,19 rather than the typical polyadenylation signal, AATAAA. To identify a consensus sequence of the signals in red algal lineage, accumulation of a lot of cDNA data, including 3′ UTR, would be required.…”

Section: Discussionmentioning

confidence: 97%

“…Porphyra yezoensis is known as the most economically valuable seaweed in Japan, and it has recently been recognized as a model laboratory organism for fundamental and applied studies in marine sciences because of easy cultivation in a small space, short generation time and a small genome. 1,2 In addition, developments of transformation systems, using both the alga itself [3][4][5] and yeast (Endo et al unpubl. data, 2006) as a host cell, are in progress to perform functional analysis of genes of the alga.…”

Section: Introductionmentioning

confidence: 99%

Characterization of a cDNA encoding a homolog of actin-related protein 4 from a marine red alga Porphyra yezoensis

et al. 2006

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A cDNA (PyARP4) containing an open reading frame for a protein of 573 amino acids was identified in the marine red alga Porphyra yezoensis. The conceptual PyARP4 protein exhibits significant similarity to actin-related protein (ARP) 4 in the terrestrial plant Arabidopsis. Comparison of the deduced amino acid sequence showed moderate sequence identity (30%) to a conventional actin in P. yezoensis, as seen in comparisons between ARP and conventional actins of other organisms. A putative bipartite nuclear localization signal and an actin motif were found within the PyARP4 amino acid sequence. In a phylogenetic analysis, the PyARP4 was found to cluster with the ARP4 of other organisms. The expression level of PyARP4 did not change significantly among four developmental stages of life cycle and was lower than that of a conventional actin. This cDNA therefore may serve as a useful internal standard in gene expression analyses of differentially expressed genes in P. yezoensis.

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“…Five primer pairs were designed based on nucleotide sequences of ß-tubulin, TOP2, EF-1 including 5 0 -untranslated region (UTR) and V-ATPase from P. yezoensis, as reported in previous studies (Kitade et al, 1999;Shimomura et al, 2002;Fukuda et al, 2003;Gong et al, 2005, Table 2). To evaluate the influence of the presence of introns on the level of polymorphism, we used three primer pairs that were not expected to span an intron for amplification, i.e.…”

Section: Caps Analysis: Pcr Amplification and Restriction Endonucleasmentioning

confidence: 99%

Genetic polymorphism withinPorphyra yezoensis(Bangiales, Rhodophyta) and related species from Japan and Korea detected by cleaved amplified polymorphic sequence analysis

Park

Fukuda

Endo

et al. 2007

European Journal of Phycology

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We investigated genetic diversity in 10 strains of Porphyra yezoensis and related species, collected from Japan and Korea, in order to identify an appropriate crossing partner for use in various genetic analyses. Firstly, the phylogenetic relationships among the 10 strains were examined using SSU rDNA sequences. All five Japanese strains (TU-1, TU-2, TUH-25, JHS and JHU) and two of the Korean strains (KGJ and KPH) were identified as P. yezoensis while the other three Korean strains (KTY1, KTY2 and KTY3) were identified as P. tenera. Using cleaved amplified polymorphic sequence analysis, genetic polymorphisms lying in five regions of four genes (ß-tubulin, TOP2, EF-1 and V-ATPase) were examined. Twenty out of 34 restriction endonucleases revealed genetic polymorphisms between the seven strains of P. yezoensis and the three strains of P. tenera in all five regions tested. Furthermore, when they were employed to digest the fragments of TOP2 and V-ATPase, 14 enzymes discriminated the two Korean strains of P. yezoensis (KGJ and KPH) from the five Japanese strains of this species. The level of polymorphism was much higher in the amplified fragment of V-ATPase (including four introns) than in that of TOP2 without an intron. The gene regions with detected polymorphisms would be useful molecular markers for confirmation of cross-fertilization in sporophytic thalli and for construction of a linkage map. A suitable combination of the strains in the cross experiment is discussed.

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Isolation, characterization and expression of a cDNA encoding an elongation factor-1alpha from Porphyra yezoensis (Bangiales, Rhodophyta)

Cited by 5 publications

References 19 publications

Variation of Expression Levels of Seven Housekeeping Genes at Different Life-History Stages in Porphyra yezoensis

Variation of Expression Levels of Seven Housekeeping Genes at Different Life-History Stages in Porphyra yezoensis

Characterization of a cDNA encoding a homolog of actin-related protein 4 from a marine red alga Porphyra yezoensis

Genetic polymorphism withinPorphyra yezoensis(Bangiales, Rhodophyta) and related species from Japan and Korea detected by cleaved amplified polymorphic sequence analysis

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