MOLECULAR IDENTIFICATION THROUGH 16S rRNA GENE SEQUENCING OF PARAMAGNETOSPIRILLUM MAGNETICUM AMB-1 FROM WARSAK, PAKISTAN
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Keywords
Bacteria, Extraction, Culturing, PCR, Gene sequencing
Abstract
Paramagnetospirillum magneticum AMB-1 is a gram-negative, facultative anaerobic bacterium. The aim of current study is molecular identification through 16S rRNA gene sequencing of magnetotactic bacteria (Paramagnetospirillum magneticum AMB-1) from freshwater sample. Water sample was collected from the edge of freshwater site near warsak dam and magnets were used for further isolation of magnetotactic bacteria. The ATCC medium 1653 was used for culturing the bacteria. The selected bacterial strain whole genomic DNA was extracted using the phenol/chloroform DNA extraction (SDS-based technique) which was then amplified using PCR. The strain's 16S rRNA sequencing was done by the Centre for Genomic Sciences (CGS). MEGA 11.0 software is then used to perform phylogenetic analysis on these sequences and the other similar sequences. The bacterial culture showed whitish colour and spherical shape. The elevation and edge of culture was convex and entire. Subject's DNA was amplified and aligned onto a molecular ladder, as confirmed by the PCR results, which were visible at 1500 bp DNA molecular ladder. The amplification efficiency of polymerase chain reaction was very almost 100% which is generally considered the best parameter study. The isolated AMB-1 strain of bacteria exhibited 99% and 81% similarity to the magnetotactic bacteria Magnetospirllum magneticum and Magnetospirllum sp., respectively. The current research contributed to the identification of this bacterial isolate which was discovered for the 1st time in Pakistan’s freshwater sample.
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Amna Nisar
Department of Pharmacy, University of Peshawar, Peshawar 25120, Khyber Pakhtunkhwa, Pakistan;
Shumaila Rauf
Department of Pharmacy, University of Peshawar, Peshawar 25120, Khyber Pakhtunkhwa, Pakistan;
Fazle Rabbi
Department of Pharmacy, Abasyn University Peshawar, Peshawar 25000, Khyber Pakhtunkhwa, Pakistan;