Isolasi dan Pengklonan Fragmen eDNA Cen Penyandi H+-ATPase Membran Plasma dari Melastoma malabathricum L.

Abstract

Melastoma malabathricum L. grows well in acid soil with high level a/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 Ihis research was 10 isolale and clone Ihe cDNA fragmenl of MmPMA encoding plasma membrane H~ -ATPase from M. malabalhricum L. By reverse Iranscriplion. 10101 cDNA had been synthesized from Ihe 10101 RNA as lemplate. The fragmenl of MmPMA cDNA had been successfully isolaled by PCR by using 10101 cDNA as lemplale and PMA primer designed from conserved region Jor corresponding gene. This fragment had been success/ully inserted into pGEM-T Easy and the recombinant plasmid was sl/cces!J!ully introduced into ft.. coli DH5a. Nucleotide sequ.ence analysis showed that the length of MmPMA frogmen I is 806 bp encoding 268 amino acids. Local alignment analysis based on nucleotide of mRNA showed Ihal MmPMA fragmenl was 81% idenlical 10 pari of PMA of Sesbania roslrala Juglam regia and Prunus persico. Based on deduced amino acid sequence, MmPMA was 94% idenlicallo pari of PMA of Juglam regia; 93% 10 PMA ofS. roslrala, and Arabidopsis Iltaliana. MmPMAfragmenl has pltosphO/ylalion inlermediale domain (DKTGT) and ATP binding domain (KGAp' DPPR. MITGD, andGDGVN).