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Utut Widyastuti Suharsono
Departemen Biologi, Fakultas Matematika dan Ilmu Pengetahuan Alam, Institut Pertanian Bogor (Bogor Agricultural University), Kampus IPB Darmaga, Bogor 16680, Indonesia

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ISOLASI DAN PENGKLONAN FRAGMEN cDNA DARI GEN PENYANDI MULTIDRUG RESISTANCE ASSOCIATED PROTEIN DARI Melastoma affine Suharsono, Suharsono; Firdaus, Syarifin; Suharsono, Utut Widyastuti
Makara Journal of Science Vol. 12, No. 2
Publisher : UI Scholars Hub

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Isolation and Cloning of cDNA Fragment of Gene Encoding for Multidrug Resistance Associated Protein from M. affine. M. affine can grow well in acid soil with high level of soluble aluminum. One of the important proteins in the detoxifying xenobiotic stress including acid and Al stresses is a multidrug resistance associated protein (MRP) encoded by mrp gene. The objective of this research is to isolate and clone the cDNA fragment of MaMrp encoding MRP from M. affine. By reverse transcription, total cDNA had been synthesized from the total RNA as template. The fragment of cDNA MaMrp had been successfully isolated by PCR by using total cDNA as template and mrp primer designed from A. thaliana, yeast, and human. This fragment was successfully inserted into pGEM-T Easy and the recombinant plasmid was successfully introduced into E. coli DH5α. Nucleotide sequence analysis showed that the lenght of MaMrp fragment is 633 bp encoding 208 amino acids. Local alignment analysis based on nucleotide of mRNA showed that MaMrp fragment is 69% identical to AtMrp1 and 63% to AtMrp from A. thaliana. Based on deduced amino acid sequence, MaMRP is 84% identical to part of AtMRP13, 77% to AtMRP12, and 73% to AtMRP1 from A. thaliana respectively. Alignment analysis with AtMRP1 showed that MaMRP fragment is located in TM1 and NBF1 domains and has a specific amino acid sequence QCKAQLQNMEEE
ISOLASI DAN PENGKLONAN FRAGMEN cDNA DARI GEN PENYANDI MULTIDRUG RESISTANCE ASSOCIATED PROTEIN DARI Melastoma affine Suharsono, Suharsono; Firdaus, Syarifin; Suharsono, Utut Widyastuti
Makara Journal of Science Vol. 11, No. 2
Publisher : UI Scholars Hub

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Isolation and Cloning of cDNA Fragment of Gene Encoding for Multidrug Resistance Associated Protein from M. affine. M. affine can grow well in acid soil with high level of soluble aluminum. One of the important proteins in the detoxifying xenobiotic stress including acid and Al stresses is a multidrug resistance associated protein (MRP) encoded by mrp gene. The objective of this research is to isolate and clone the cDNA fragment of MaMrp encoding MRP from M. affine. By reverse transcription, total cDNA had been synthesized from the total RNA as template. The fragment of cDNA MaMrp had been successfully isolated by PCR by using total cDNA as template and mrp primer designed from A. thaliana, yeast, and human. This fragment was successfully inserted into pGEM-T Easy and the recombinant plasmid was successfully introduced into E. coli DH5α. Nucleotide sequence analysis showed that the lenght of MaMrp fragment is 633 bp encoding 208 amino acids. Local alignment analysis based on nucleotide of mRNA showed that MaMrp fragment is 69% identical to AtMrp1 and 63% to AtMrp from A. thaliana. Based on deduced amino acid sequence, MaMRP is 84% identical to part of AtMRP13, 77% to AtMRP12, and 73% to AtMRP1 from A. thaliana respectively. Alignment analysis with AtMRP1 showed that MaMRP fragment is located in TM1 and NBF1 domains and has a specific amino acid sequence QCKAQLQNMEEE
ISOLASI FRAGMEN cDNA DARI GEN PENYANDI AKTIN DARI Melastoma malabathricum Hannum, Saleha; Akashi, Kinya; Suharsono, Utut Widyastuti; Hartana, Alex
Makara Journal of Science Vol. 14, No. 2
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Isolation of cDNA Fragment of Gene Encoding for Actin from Melastoma malabthricum. M. malabathricum grows well in acidic soil with high Al solubility, thereby it can be used as a model plant for tolerance to aluminum and acid stresses. Actin is housekeeping gene used as an internal control for gene expression analysis. The objective of this research was to isolate and clone the cDNA fragments of MmACT encoding for actin of M. malabathricum. Total RNA was isolated and used as the template for cDNA synthesis by reverse transcription. Four cDNA fragments of MmACT, called MmACT1, MmACT2, MmACT3, and MmACT4, had been isolated and inserted into pGEM-T Easy plasmid. Nucleotide sequence analysis showed that the size of MmACT1 and MmACT2 is 617 bp, whereas MmACT3 and MmACT4 is 735 bp. The similarity among these four MmACT is about 78%-99% based on nucleotide sequence and about 98%-100% based on amino acid sequence. Phylogenetic analysis based on amino acid sequence showed that at 1% dissimilarity, the MmACT1, MmACT2, MmACT3 and the ACT5 Populus trichocarpha are clustered in one group, while the MmACT4 is grouped with ACT9 P. trichocarpa and ACT1 Gossypium hirsutum, and these two groups are separated from actin group of monocotyledonous plants. The sequence of MmACT fragments were registered in GenBank/EMBL/DDBJ database with accession numbers AB500686, AB500687, AB500688, and AB500689.
ISOLASI DAN PENGKLONAN FRAGMEN cDNA DARI GEN PENYANDI MULTIDRUG RESISTANCE ASSOCIATED PROTEIN DARI Melastoma affine Suharsono, Suharsono; Firdaus, Syarifin; Suharsono, Utut Widyastuti
Makara Journal of Science Vol. 12, No. 2
Publisher : UI Scholars Hub

Show Abstract | Download Original | Original Source | Check in Google Scholar

Abstract

Isolation and Cloning of cDNA Fragment of Gene Encoding for Multidrug Resistance Associated Protein from M. affine. M. affine can grow well in acid soil with high level of soluble aluminum. One of the important proteins in the detoxifying xenobiotic stress including acid and Al stresses is a multidrug resistance associated protein (MRP) encoded by mrp gene. The objective of this research is to isolate and clone the cDNA fragment of MaMrp encoding MRP from M. affine. By reverse transcription, total cDNA had been synthesized from the total RNA as template. The fragment of cDNA MaMrp had been successfully isolated by PCR by using total cDNA as template and mrp primer designed from A. thaliana, yeast, and human. This fragment was successfully inserted into pGEM-T Easy and the recombinant plasmid was successfully introduced into E. coli DH5α. Nucleotide sequence analysis showed that the lenght of MaMrp fragment is 633 bp encoding 208 amino acids. Local alignment analysis based on nucleotide of mRNA showed that MaMrp fragment is 69% identical to AtMrp1 and 63% to AtMrp from A. thaliana. Based on deduced amino acid sequence, MaMRP is 84% identical to part of AtMRP13, 77% to AtMRP12, and 73% to AtMRP1 from A. thaliana respectively. Alignment analysis with AtMRP1 showed that MaMRP fragment is located in TM1 and NBF1 domains and has a specific amino acid sequence QCKAQLQNMEEE
Co-Authors ALEX HARTANA Firdaus, Syarifin KINYA AKASHI Saleha Hannum Suharsono Suharsono