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ISOLATION AND CLONING OF cDNA OF GENE ENCODING FOR METALLOTHIONEIN TYPE 2 FROM MELASTOMA AFFINE

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Author(s): SUHARSONO | NIKEN TRISNANINGRUM | LULUT DWI SULISTYANINGSIH | UTUT WIDYASTUT

Journal: BIOTROPIA : the Southeast Asian Journal of Tropical Biology
ISSN 0215-6334

Volume: 16;
Issue: 1;
Start page: 28;
Date: 2009;
Original page

Keywords: cDNA | metallothionein | Melastoma affine | cloning | cysteine

ABSTRACT
Metallothionein is an important protein for detoxifying heavy metal ions. h is research was conducted to isolate and clone cDNA of gene encoding for metallothionein type 2 from Melastoma affi ne . Total RNA was isolated from young leaves. Total cDNA was synthesized from the total RNA by reverse transcription. h e MaMt2 cDNA was successfully isolated by PCR technique. h e MaMt2 cDNA was inserted into pGEM-T Easy and the recombinant plasmid was successfully introduced into Escherichia coli DH5 α . DNA sequencing analysis showed that this cDNA is full length consisting of 246 pb encoding 81 amino acid residues. h is cDNA is identical to mRNA of AtMt2 from Arabidopsis thaliana. It does not contain any restriction sites found in the cloning sites of pGEM-T Easy. h e deduced protein of MaMT2 contains 14 cysteine residues distributed in the Cys-Cys, Cys-X-Cys, and Cys-X-X-Cys motifs
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