Helvin, Vincent; Saritha G, Bhatt(Cochin University of Science and Technology, May 7, 2014)
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Abstract:
Soil community genomics or metagenomics is employed in this study to analyze the evolutionary related
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ness of mangrove microbial community. The metagenomic DNA was isolated from mangrove sediment and
16SrDNA was amplified using universal primers. The amplicons were ligated into pTZ57R/T cloning vector and transformed
onto E. coli JM109 host cells. The recombinant plasmids were isolated from positive clones and the insert was confirmed
by its reamplification. The amplicons were subjected to Amplified Ribosomal DNA Restriction Analysis (ARDRA) using three
different tetra cutter restriction enzymes namely Sau3A1, Hha1 and HpaII. The 16SrDNA insert were sequenced and their
identity was determined. The sequences were submitted to NCBI database and accession numbers obtained. The phylo
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genetic tree was constructed based on Neighbor-Joining technique. Clones belonged to two major phyla of the bacterial
domain, namely Firmicutes and Proteobacteria, with members of Firmicutes predominating. The microbial diversity of the
mangrove sediment was explored in this manner.