ISOLASI FRAGMEN cDNA DARI GEN PENYANDI AKTIN DARI Melastoma malabathricum

Hannum, Saleha; Akashi, Kinya; Suharsono, Utut Widyastuti; Hartana, Alex; Yokota, Akiho; Suharsono, _

doi:10.7454/mss.v14i2.730

Cited by 7 publications

(11 citation statements)

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“…clone Riau can then be designed in both exon regions based on the obtained sequence. Such primer designing -namely P_act_F: 5'-TGG TTG GCA TGG GTC AGA A-3' and P_act_R: 5'-GTG ATG ACT TGG CCG TCT GG-3' -is aimed for certain genes ex pression studies necessity as well as a genomic 256 Volume 9 | Number 3 | September | 2019 DNA contamination verification on a target cDNA [5,6,15]. For this purpose, amplification is conducted using DNA and cDNA molecules separately.…”

Section: Resultsmentioning

confidence: 99%

“…Four members of Melastoma malabathricum actin genes family, i.e. MmACT1, MmACT2, MmACT3, and MmACT4 [5] and 10 of Arabidopsis thaliana ones [6] have also been identified and characterized. One of A. thaliana actin genes, such as AtACT2, is most frequent and more accurate to be applied as an internal control in A. thaliana gene expression analysis [7].…”

Section: Introductionmentioning

confidence: 99%

“…Four degenerate actin primers have been designed based on actin gene conserved regions among the known plant, animal, fungal, and protist, namely PlAc12S and PlAc46S-20 as forward primers and PlAc245N-20 and PlAc284N as reverse primers [6]. The four primers have been applied in Melastoma malabathricum [5]. A primer pair of PlAc12S-20 and PlAc245N-20 has also been used to isolate E. floribundus actin gene [13].…”

Section: Introductionmentioning

confidence: 99%

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The DNA Sequence Encoding Actin (ACT1) of Pandan (Benstonea sp.)

Wahyuningsih

Elyani²,

Damayanti³

et al. 2019

J.Trop.Life.Science

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One of housekeeping genes is actin gene. This gene is frequently used in gene expression studies as an internal control. The DNA sequence encoding actin from Pandan (Benstonea sp.) clone Riau has not been reported, therefore, this study investigated the DNA sequence encoding actin isolated from Benstonea sp. clone Riau. Total DNA isolation was performed in fresh leaves, total RNA isolation from stem, total cDNA synthesis, polymerase chain reaction using degenerate actin primer, electrophoresis, cloning, transformation, blue white colony selection, colony PCR, sequencing, data analysis using BioEdit and MEGA6 softwares and BLASTn program. The partial DNA sequence encoding actin from Benstonea sp. clone Riau obtained was 1,403 bp. The sequence was grouped as part of actin1 (ACT1) and it was consisted of two exons and one intron. The predicted coding and peptide sequences were 616 bp and 205 amino acids, respectively. The predicted coding sequence had 90% similarity to some ACT1 mRNA from some plants species but none of which belongs to Benstonea genus or Pandanaceae family. The deduced peptide sequence had similarity to some ACT1 peptide from some plant species of up to 99% and also none of them belongs to Benstonea genus or Pandanaceae family. Thus, the partial ACT1 gene obtained in this study was the first sequence reported from Benstonea genus. Furthermore, this sequence can be used as a reference to isolate actin genes from other species within Benstonea genus for gene expression analysis purposes.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

The DNA Sequence Encoding Actin (ACT1) of Pandan (Benstonea sp.)

Wahyuningsih

Elyani²,

Damayanti³

et al. 2019

J.Trop.Life.Science

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“…Such conditions cause the EF1α to be able to be used as a marker to determine DNA contamination in total cDNA in addition to its potential as internal control in gene expression studies. For this purpose, PCR using DNA or cDNA molecule as template was performed separately and after that the sizes of the PCR products from the two templates were compared (Hannum et al, 2010). In this case, the PCR product from DNA template around 850 bp in length and cDNA template will produce a shorter PCR product which is about 750 bp in length.…”

Section: Analysis Of Partial Housekeeping Gene Sequencesmentioning

confidence: 99%

Isolation of Partial Housekeeping Genes on Tuntun Angin (Elaeocarpus floribundu)

et al. 2019

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Some genes like 18S rRNA, 26S rRNA, elongation factor 1-alpha (EF1α), and beta-tubulin (TUB) are members of housekeeping genes group that are commonly used as internal control in gene expression study. This study aimed to isolate those four housekeeping genes of tuntun angin (Elaeocarpus floribundus). The research material included fresh leaves of E. floribundus that were picked up from Kajuik Lake in Riau Province and four primer pairs. The procedures consisted of total DNA isolation using Genomic DNA Mini Kit Plant (Geneaid), polymerase chain reaction (PCR), electrophoresis on 1% agarose gel, sequencing, and bioinformatic analysis. This study has been isolated 18S rRNA, 26S rRNA, EF1α, and TUB genes with the size of 422 bp, 922 bp, 856 bp, and 877 bp, respectively. The EF1α and TUB genes has never been reported in Elaeocarpaceae family. Thus, those partial DNA sequences are the first sequences reported from this species and can be used as reference genes in this plant after validation.

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“…Some housekeeping genes have been isolated from various plants, such as the actin from the plant of Arabidopsis thaliana (McDowell et al, 1996), Melastoma malabathricum (Hannum et al, 2010), Oryza sativa (McElroy et al, 1990), Zea mays (Staiger et al, 2000), Jatropha curcas (Yuniati et al, 2011) and Elaeocarpus floribundus (Roslim & Herman, 2017a). The tubulin-coding gene has been isolated from the Arabidopsis thaliana plant (Snustad et al, 1992) and Linum usitatissimum (Gavazzi et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

Isolation of Housekeeping Genes on Durik-durik (Syzygium sp)

et al. 2018

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Housekeeping gene is a gene expressed with a fixed level and in abundant amounts under various conditions. After validation, the housekeeping gene can be used as an internal control to normalize gene expression data. This study reports the isolation of several housekeeping genes in Durik-durik plant (Syzygium sp). This plant material in form of fresh leaves from Durik-durik plants are taken from Kajuik Lake, Riau Province. The next stage is total DNA isolation, polymerase chain reaction, electrophoresis, sequencing and data analysis using bioinformatic tools. The isolated housekeeping genes included 18S rRNA, actin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), Beta-tubulin and Ubiquitin with sizes of 400 bp, 679 bp, 1134 bp, 836 bp, 1167 bp and 2155 bp, respectively. In addition to 18S rRNA, the five housekeeping genes are the first reported from the genus Syzygium and referable to isolate housekeeping genes in other species in this genus. The six housekeeping genes can be used as internal controls on Durik-durik plants after validation.

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ISOLASI FRAGMEN cDNA DARI GEN PENYANDI AKTIN DARI Melastoma malabathricum

Cited by 7 publications

References 0 publications

The DNA Sequence Encoding Actin (ACT1) of Pandan (Benstonea sp.)

The DNA Sequence Encoding Actin (ACT1) of Pandan (Benstonea sp.)

Isolation of Partial Housekeeping Genes on Tuntun Angin (Elaeocarpus floribundu)

Isolation of Housekeeping Genes on Durik-durik (Syzygium sp)

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