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Biochemical & Molecular Characterization Of Locally Isolated Extremophile

By: Iram Murtaza | Dr. Muhammad Tayyab.
Contributor(s): Ms. Sehrish | Ms. Shagufta Saeed.
Material type: materialTypeLabelBookPublisher: 2013Subject(s): Institute of Biochemistry & BiotechnologyDDC classification: 1706,T Dissertation note: Extremophiles are microorganisms with the ability to survive under extreme of conditions. Due to their extreme stability, these microorganisms produce unique biocatalysts that have been exploited in various industrial processes. These micro-organisms are unique factories for the production of enzymes that have great potential for agriculture, textile, pharmaceutical, poultry and detergent industries. The present study was conducted for the isolation and characterization of alkaliphile. The sampling was done from spring located in Rawat, Pakistan. Optimization of growth conditions was done by growing the microorganism at various conditions including temperature, pH and salt concentration. The microorganism was identified on the basis of biochemical characteristics as well as on the basis of 16S rRNA gene sequence. Regarding the molecular characterization, the genomic DNA was isolated from the strain and was utilized for the amplification of 16S rRNA gene. The PCR product was ligated in pTZ57R/T. The ligation mixture was utilized for the transformation of E.coli DH5-? cells. The presence of insert in recombinant pTZ57R/T was confirmed by single and double restriction with EcoR1 and Hind III which resulted in the liberation of DNA fragment. The gene sequence was utilized for the phylogenetic analysis. The microorganism was found to be Gram positive rods involved in the production of catalase, amylase, protease, enzymes and gave positive results for Mannitol, Voges Proskauer Tests while negative for citrate utilization and nitrate reduction test. 16S rRNA gene sequence analysis demonstrated that the newly isolated strain showed maximum homology with various members of genus Exiguobacterium. The newly isolated strain was declared a new member of genus Exiguobacterium and was named as Exiguobacterium UVAS-01.
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Item type Current location Collection Call number Status Date due Barcode Item holds
Thesis Thesis UVAS Library
Thesis Section
Veterinary Science 1706,T (Browse shelf) Available 1706,T
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Extremophiles are microorganisms with the ability to survive under extreme of conditions. Due to their extreme stability, these microorganisms produce unique biocatalysts that have been exploited in various industrial processes. These micro-organisms are unique factories for the production of enzymes that have great potential for agriculture, textile, pharmaceutical, poultry and detergent industries.
The present study was conducted for the isolation and characterization of alkaliphile. The sampling was done from spring located in Rawat, Pakistan. Optimization of growth conditions was done by growing the microorganism at various conditions including temperature, pH and salt concentration. The microorganism was identified on the basis of biochemical characteristics as well as on the basis of 16S rRNA gene sequence. Regarding the molecular characterization, the genomic DNA was isolated from the strain and was utilized for the amplification of 16S rRNA gene. The PCR product was ligated in pTZ57R/T. The ligation mixture was utilized for the transformation of E.coli DH5-? cells. The presence of insert in recombinant pTZ57R/T was confirmed by single and double restriction with EcoR1 and Hind III which resulted in the liberation of DNA fragment. The gene sequence was utilized for the phylogenetic analysis. The microorganism was found to be Gram positive rods involved in the production of catalase, amylase, protease, enzymes and gave positive results for Mannitol, Voges Proskauer Tests while negative for citrate utilization and nitrate reduction test. 16S rRNA gene sequence analysis demonstrated that the newly isolated strain showed maximum homology with various members of genus Exiguobacterium. The newly isolated strain was declared a new member of genus Exiguobacterium and was named as Exiguobacterium UVAS-01.

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