Screening and characterization of alpha-amylase expressing gene from amylolytic bacteria isolated from Sago humus

The amylolytic bacterial colony that yields high level of the a-amylase enzyme activity by producing the biggest halo when flooded with Lugol's Iodine into minimal starch media was isolated from sago humus. Result from the biochemical tests show that these bacteria pertain in genus of gram-posi...

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Main Author: Rohanie, Binti Bohan.
Format: Final Year Project Report / IMRAD
Language:English
Published: Universiti Malaysia Sarawak (UNIMAS) 2006
Subjects:
Online Access:http://ir.unimas.my/id/eprint/18948/
http://ir.unimas.my/id/eprint/18948/1/Rohanie%20ft.pdf
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author Rohanie, Binti Bohan.
author_facet Rohanie, Binti Bohan.
author_sort Rohanie, Binti Bohan.
building UNIMAS Institutional Repository
collection Online Access
description The amylolytic bacterial colony that yields high level of the a-amylase enzyme activity by producing the biggest halo when flooded with Lugol's Iodine into minimal starch media was isolated from sago humus. Result from the biochemical tests show that these bacteria pertain in genus of gram-positive Bacillus spp. However, the molecular identification of the isolated bacteria by direct sequencing of 16S rRNA purified PCR products was unsuccessful. In order to isolate a-amylase gene from the isolated bacteria, Polymerase Chain Reaction (PCR) technique using two different set of specific primers namely ApAl&ApA2 and AmBcl&AmBc2 primers was done. Approximately 500bp and 750bp a-amylase gene were successfully isolated respectively using both sets of primers which then purified and sent for direct sequencing. These PCR products were also cloned into pGEM-T Easy vector system (Promega) in Escherichia coli XL-I Blue competent cells for propagation of the products. The white colonies were then screened for insert using colony PCR and Restriction Endonuclease analysis.
first_indexed 2025-11-15T06:58:23Z
format Final Year Project Report / IMRAD
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institution Universiti Malaysia Sarawak
institution_category Local University
language English
last_indexed 2025-11-15T06:58:23Z
publishDate 2006
publisher Universiti Malaysia Sarawak (UNIMAS)
recordtype eprints
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spelling unimas-189482024-06-25T07:42:30Z http://ir.unimas.my/id/eprint/18948/ Screening and characterization of alpha-amylase expressing gene from amylolytic bacteria isolated from Sago humus Rohanie, Binti Bohan. QR Microbiology The amylolytic bacterial colony that yields high level of the a-amylase enzyme activity by producing the biggest halo when flooded with Lugol's Iodine into minimal starch media was isolated from sago humus. Result from the biochemical tests show that these bacteria pertain in genus of gram-positive Bacillus spp. However, the molecular identification of the isolated bacteria by direct sequencing of 16S rRNA purified PCR products was unsuccessful. In order to isolate a-amylase gene from the isolated bacteria, Polymerase Chain Reaction (PCR) technique using two different set of specific primers namely ApAl&ApA2 and AmBcl&AmBc2 primers was done. Approximately 500bp and 750bp a-amylase gene were successfully isolated respectively using both sets of primers which then purified and sent for direct sequencing. These PCR products were also cloned into pGEM-T Easy vector system (Promega) in Escherichia coli XL-I Blue competent cells for propagation of the products. The white colonies were then screened for insert using colony PCR and Restriction Endonuclease analysis. Universiti Malaysia Sarawak (UNIMAS) 2006 Final Year Project Report / IMRAD NonPeerReviewed text en http://ir.unimas.my/id/eprint/18948/1/Rohanie%20ft.pdf Rohanie, Binti Bohan. (2006) Screening and characterization of alpha-amylase expressing gene from amylolytic bacteria isolated from Sago humus. [Final Year Project Report / IMRAD] (Unpublished)
spellingShingle QR Microbiology
Rohanie, Binti Bohan.
Screening and characterization of alpha-amylase expressing gene from amylolytic bacteria isolated from Sago humus
title Screening and characterization of alpha-amylase expressing gene from amylolytic bacteria isolated from Sago humus
title_full Screening and characterization of alpha-amylase expressing gene from amylolytic bacteria isolated from Sago humus
title_fullStr Screening and characterization of alpha-amylase expressing gene from amylolytic bacteria isolated from Sago humus
title_full_unstemmed Screening and characterization of alpha-amylase expressing gene from amylolytic bacteria isolated from Sago humus
title_short Screening and characterization of alpha-amylase expressing gene from amylolytic bacteria isolated from Sago humus
title_sort screening and characterization of alpha-amylase expressing gene from amylolytic bacteria isolated from sago humus
topic QR Microbiology
url http://ir.unimas.my/id/eprint/18948/
http://ir.unimas.my/id/eprint/18948/1/Rohanie%20ft.pdf