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All Journal Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy)
Utut Widyastuti Suharsono
Departemen Biologi, Fakultas Matematika dan Ilmu Pengetahuan Alam, Institut Pertanian Bogor (Bogor Agricultural University),
Agriculture, Biological Sciences & Forestry Economics, Econometrics & Finance

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Isolasi dan Pengklonan Fragmen cDNA Gen Penyandi H+-ATPase Membran Plasma dari Melastoma malabathricum L. , Muzuni; Didy Sopandie; Utut Widyastuti Suharsono; , Suharsono
Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy) Vol. 38 No. 1 (2010): Jurnal Agronomi Indonesia
Publisher : Indonesia Society of Agronomy (PERAGI) and Department of Agronomy and Horticulture, Faculty of Agriculture, IPB University, Bogor, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (378.981 KB) | DOI: 10.24831/jai.v38i1.1680

Abstract

Melastoma malabathricum L. grows well in acid soil with high level of soluble aluminum. One of the important proteins in the detoxifying acid and aluminum stress is a plasma membrane H+  -ATPase protein encoded by PMA gene. The objective of this research was to isolate and clone the cDNA fragment of MmPMA encoding plasma membrane H+ -ATPase from M. malabathricum L. By reverse transcription, total cDNA had been synthesized from the total RNA as template. The fragment of MmPMA  cDNA  had been successfully isolated by PCR by using total cDNA  as  template and PMA primer designed from conserved region for corresponding gene. This fragment had been successfully inserted into pGEM-T Easy and the recombinant plasmid was successfully introduced into E. coli DH5". Nucleotide sequence analysis showed that the length of MmPMA fragment is 806 bp encoding 268 amino acids. Local alignment analysis based on nucleotide of mRNA showed that MmPMA fragment was 81% identical to part of PMA of Sesbania rostrata, Juglans regia, and Prunus persica. Based on deduced amino acid sequence, MmPMA was 94% identical to part of PMA of Juglans regia; 93% to PMA of S. rostrata, and Arabidopsis thaliana. MmPMA fragment has phosphorylation intermediate domain (DKTGT) and ATP binding domain (KGAP, DPPR, MITGD, and GDGVN).   Keywords: isolation, Melastoma malabathricum L., MmPMA fragment, sequencing
Co-Authors , Suharsono Didy Sopandie Muzuni, Muzuni