ISOLATION AND CLONING OF cDNA OF GENE ENCODING FOR METALLOTHIONEIN TYPE 2 FROM MELASTOMA AFFINE
Metallothionein is an important protein for detoxifying heavy metal ions. This research was conducted to isolate and clone cDNA of gene encoding for metallothionein type 2 from Melastoma affine. Total RNA was isolated from young leaves. Total cDNA was synthesized from the total RNA by reverse tra...
Main Authors: | , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
SEAMEO, BIOTROP
2009-06-01
|
Series: | Biotropia: The Southeast Asian Journal of Tropical Biology |
Online Access: | https://journal.biotrop.org/index.php/biotropia/article/view/64 |
_version_ | 1797758589521100800 |
---|---|
author | SUHARSONO SUHARSONO NIKEN TRISNANINGRUM LULUT DWI SULISTYANINGSIH UTUT WIDYASTUTI |
author_facet | SUHARSONO SUHARSONO NIKEN TRISNANINGRUM LULUT DWI SULISTYANINGSIH UTUT WIDYASTUTI |
author_sort | SUHARSONO SUHARSONO |
collection | DOAJ |
description |
Metallothionein is an important protein for detoxifying heavy metal ions. This research was conducted to isolate and clone cDNA of gene encoding for metallothionein type 2 from Melastoma affine. Total RNA was isolated from young leaves. Total cDNA was synthesized from the total RNA by reverse transcription. The MaMt2 cDNA was successfully isolated by PCR technique. The MaMt2 cDNA was inserted into pGEM-T Easy and the recombinant plasmid was successfully introduced into Escherichia coli DH5α. DNA sequencing analysis showed that this cDNA is full length consisting of 246 pb encoding 81 amino acid residues. This 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. The deduced protein of MaMT2 contains 14 cysteine residues distributed in the Cys-Cys, Cys-X-Cys, and Cys-X-X-Cys motifs.
Â
Key words: cDNA, metallothionein, Melastoma affine, cloning, cysteine
|
first_indexed | 2024-03-12T18:32:11Z |
format | Article |
id | doaj.art-b39f5c2131a6440bb61d7e1b5602acbb |
institution | Directory Open Access Journal |
issn | 0215-6334 1907-770X |
language | English |
last_indexed | 2024-03-12T18:32:11Z |
publishDate | 2009-06-01 |
publisher | SEAMEO, BIOTROP |
record_format | Article |
series | Biotropia: The Southeast Asian Journal of Tropical Biology |
spelling | doaj.art-b39f5c2131a6440bb61d7e1b5602acbb2023-08-02T08:17:12ZengSEAMEO, BIOTROPBiotropia: The Southeast Asian Journal of Tropical Biology0215-63341907-770X2009-06-0116110.11598/btb.2009.16.1.64ISOLATION AND CLONING OF cDNA OF GENE ENCODING FOR METALLOTHIONEIN TYPE 2 FROM MELASTOMA AFFINESUHARSONO SUHARSONO0NIKEN TRISNANINGRUM1LULUT DWI SULISTYANINGSIH2UTUT WIDYASTUTI31 Research Center for Bioresources and Biotechnology1Research Center for Bioresources and Biotechnology1Research Center for Bioresources and Biotechnology1Research Center for Bioresources and Biotechnology, and 2Department of Biology, Bogor Agricultural University Indonesia Metallothionein is an important protein for detoxifying heavy metal ions. This research was conducted to isolate and clone cDNA of gene encoding for metallothionein type 2 from Melastoma affine. Total RNA was isolated from young leaves. Total cDNA was synthesized from the total RNA by reverse transcription. The MaMt2 cDNA was successfully isolated by PCR technique. The MaMt2 cDNA was inserted into pGEM-T Easy and the recombinant plasmid was successfully introduced into Escherichia coli DH5α. DNA sequencing analysis showed that this cDNA is full length consisting of 246 pb encoding 81 amino acid residues. This 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. The deduced protein of MaMT2 contains 14 cysteine residues distributed in the Cys-Cys, Cys-X-Cys, and Cys-X-X-Cys motifs.  Key words: cDNA, metallothionein, Melastoma affine, cloning, cysteine https://journal.biotrop.org/index.php/biotropia/article/view/64 |
spellingShingle | SUHARSONO SUHARSONO NIKEN TRISNANINGRUM LULUT DWI SULISTYANINGSIH UTUT WIDYASTUTI ISOLATION AND CLONING OF cDNA OF GENE ENCODING FOR METALLOTHIONEIN TYPE 2 FROM MELASTOMA AFFINE Biotropia: The Southeast Asian Journal of Tropical Biology |
title | ISOLATION AND CLONING OF cDNA OF GENE ENCODING FOR METALLOTHIONEIN TYPE 2 FROM MELASTOMA AFFINE |
title_full | ISOLATION AND CLONING OF cDNA OF GENE ENCODING FOR METALLOTHIONEIN TYPE 2 FROM MELASTOMA AFFINE |
title_fullStr | ISOLATION AND CLONING OF cDNA OF GENE ENCODING FOR METALLOTHIONEIN TYPE 2 FROM MELASTOMA AFFINE |
title_full_unstemmed | ISOLATION AND CLONING OF cDNA OF GENE ENCODING FOR METALLOTHIONEIN TYPE 2 FROM MELASTOMA AFFINE |
title_short | ISOLATION AND CLONING OF cDNA OF GENE ENCODING FOR METALLOTHIONEIN TYPE 2 FROM MELASTOMA AFFINE |
title_sort | isolation and cloning of cdna of gene encoding for metallothionein type 2 from melastoma affine |
url | https://journal.biotrop.org/index.php/biotropia/article/view/64 |
work_keys_str_mv | AT suharsonosuharsono isolationandcloningofcdnaofgeneencodingformetallothioneintype2frommelastomaaffine AT nikentrisnaningrum isolationandcloningofcdnaofgeneencodingformetallothioneintype2frommelastomaaffine AT lulutdwisulistyaningsih isolationandcloningofcdnaofgeneencodingformetallothioneintype2frommelastomaaffine AT ututwidyastuti isolationandcloningofcdnaofgeneencodingformetallothioneintype2frommelastomaaffine |