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Mangrove ecosystems constitute about 60 – 70% of costline of earth. There are many factors which make mangroves hotspots of microbial diversity. And these communities play an important role in maintenance of this ecosystem. The countless roles of these microbes in this ecosystem is appreciated but the inability to culture microbes in any environmental sample completely is also realized. Hence, this study was undertaken for accomplishing the potentials of environmental DNA of a mangrove ecosystem by constructing a metagenomic library using the isolated DNA. Metagenomic DNA (good quality, 225 μg/mL) was isolated as per standard procedure and subjected to restriction digestion using three restriction enzymes, Bam HI, Eco RI and Hind III. Plasmid with high purity and good concentration (470μg/ml) was also isolated using standard protocols. The digested fragments were ligated to a vector, pGEX 6p-1 digested with the same enzymes. The recombinant vector was used to transform the host bacterium, E.coli DH5α and a small insert metagenomic library was constructed. Transformed colonies were selected on the basis of blue-white selection. The metagenomic library was screened for amylase activity. Fifty four colonies were found to be transformed. Out of the 54 clones obtained 14 were found to show amylase activity.
Key words: Metagenomics, Environmental genomics, Metagenomic DNA, Metagenomic Library, Vector, Host, Restriction Enzymes, Restriction digestion, Clones