Community Genomics Involving Culture Independent  Approach for Assessing the Phylogenetic Diversity of  Mangrove Sediment

Helvin Vincent; Harisree P. Nair; Dr. Sarita G. Bhat

Volume : III, Issue : X, October - 2013

Community Genomics Involving Culture Independent Approach for Assessing the Phylogenetic Diversity of Mangrove Sediment

Helvin Vincent, Harisree P. Nair, Dr. Sarita G. Bhat

Abstract :

Soil community genomics or metagenomics is employed in this study to analyze the evolutionary relatedness 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 phylogenetic 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