Identification, Cloning And Expressing Of Dna Polymerase Producing Thermophile From Geothermal Water In Malaysia

Aerotolerant anaerobe, Gram-positive with long rod in shape thermophilic bacteria was successfully isolated from Ulu Legong Hot Spring, Kedah, Malaysia in this study. The thermophilic bacteria was successfully isolated from the sediment sample in this study and denoted as 3UL which showed the...

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主要作者: Hussin, Nurul Akmar
格式: Thesis
語言:English
出版: 2013
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spelling my-usm-ep.439042019-04-12T05:26:11Z Identification, Cloning And Expressing Of Dna Polymerase Producing Thermophile From Geothermal Water In Malaysia 2013-07 Hussin, Nurul Akmar QH1 Natural history (General - Including nature conservation, geographical distribution) Aerotolerant anaerobe, Gram-positive with long rod in shape thermophilic bacteria was successfully isolated from Ulu Legong Hot Spring, Kedah, Malaysia in this study. The thermophilic bacteria was successfully isolated from the sediment sample in this study and denoted as 3UL which showed the best growth and well adapted to the laboratory condition compared with other isolates at temperature ranged between 45 to 80oC. Hence, this microbe was chosen for further study. A set of universal primers (F_UNI16S and R_UNI16S) were used to amplify the 16S rRNA gene sequences for ribotyping identification methods. Its 16S rRNA gene sequences (1454 nucleotides) showed very high homology (100%) with Anoxybacillus sp. DR04. The 16S rRNA gene sequence for 3UL has been deposited into “Genbank Data Library” and assigned the accession number JQ951796. Genomic DNA from the isolate was extracted and was used to amplify DNA polymerase I gene sequences. The NREAF2 and XREAR_Fxa were forward and reverse primers used for the DNA polymerase amplification with restriction sites NcoI for forward and XhoI for reverse. The gene was 2,628 bp long and encodes a protein of 876 amino acids in length. The enzyme has molecular mass of 99 kDa and showed sequence homology with DNA polymerase I (94%) from Anoxybacillus sp., (75%) Geobacillus sp., and (74%) Bacillus sp. The gene was over expressed in Escherichia coli BL21 (DE3) by using pET28a(+) as expression vector with his-tag at the C-terminus. 2013-07 Thesis http://eprints.usm.my/43904/ http://eprints.usm.my/43904/1/Nurul%20Akmar%20Binti%20Hussin24.pdf application/pdf en public masters Universiti Sains Malaysia Pusat Pengajian Sains Kajihayat
institution Universiti Sains Malaysia
collection USM Institutional Repository
language English
topic QH1 Natural history (General - Including nature conservation
geographical distribution)
spellingShingle QH1 Natural history (General - Including nature conservation
geographical distribution)
Hussin, Nurul Akmar
Identification, Cloning And Expressing Of Dna Polymerase Producing Thermophile From Geothermal Water In Malaysia
description Aerotolerant anaerobe, Gram-positive with long rod in shape thermophilic bacteria was successfully isolated from Ulu Legong Hot Spring, Kedah, Malaysia in this study. The thermophilic bacteria was successfully isolated from the sediment sample in this study and denoted as 3UL which showed the best growth and well adapted to the laboratory condition compared with other isolates at temperature ranged between 45 to 80oC. Hence, this microbe was chosen for further study. A set of universal primers (F_UNI16S and R_UNI16S) were used to amplify the 16S rRNA gene sequences for ribotyping identification methods. Its 16S rRNA gene sequences (1454 nucleotides) showed very high homology (100%) with Anoxybacillus sp. DR04. The 16S rRNA gene sequence for 3UL has been deposited into “Genbank Data Library” and assigned the accession number JQ951796. Genomic DNA from the isolate was extracted and was used to amplify DNA polymerase I gene sequences. The NREAF2 and XREAR_Fxa were forward and reverse primers used for the DNA polymerase amplification with restriction sites NcoI for forward and XhoI for reverse. The gene was 2,628 bp long and encodes a protein of 876 amino acids in length. The enzyme has molecular mass of 99 kDa and showed sequence homology with DNA polymerase I (94%) from Anoxybacillus sp., (75%) Geobacillus sp., and (74%) Bacillus sp. The gene was over expressed in Escherichia coli BL21 (DE3) by using pET28a(+) as expression vector with his-tag at the C-terminus.
format Thesis
qualification_level Master's degree
author Hussin, Nurul Akmar
author_facet Hussin, Nurul Akmar
author_sort Hussin, Nurul Akmar
title Identification, Cloning And Expressing Of Dna Polymerase Producing Thermophile From Geothermal Water In Malaysia
title_short Identification, Cloning And Expressing Of Dna Polymerase Producing Thermophile From Geothermal Water In Malaysia
title_full Identification, Cloning And Expressing Of Dna Polymerase Producing Thermophile From Geothermal Water In Malaysia
title_fullStr Identification, Cloning And Expressing Of Dna Polymerase Producing Thermophile From Geothermal Water In Malaysia
title_full_unstemmed Identification, Cloning And Expressing Of Dna Polymerase Producing Thermophile From Geothermal Water In Malaysia
title_sort identification, cloning and expressing of dna polymerase producing thermophile from geothermal water in malaysia
granting_institution Universiti Sains Malaysia
granting_department Pusat Pengajian Sains Kajihayat
publishDate 2013
url http://eprints.usm.my/43904/1/Nurul%20Akmar%20Binti%20Hussin24.pdf
_version_ 1747821301326675968