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Microbial symbionts of sponges are well known as the sources of numerous bioactive products which are of potential industrial and pharmaceutical importance, including secondary metabolites, enzymes, lipids and heteropolysaccharides. The general inability to cultivate the majority of symbionts has hampered attempts to access their diversity and restricted exploitation of their bioactive products. However, using a recently developed, holistic molecular approach called metagenomics, the bioactive products can be accessed in a sustainable way. This approach involves construction and screening of metagenomic libraries for finding the genes of interest from uncultured symbionts, followed by transferring the genes into easily cultivated microbes, thereby producing the encoded bioactive compounds in significant amount. We report here three preliminary steps required for the shotgun based construction of metagenomic library from symbionts of a sponge 1) preparation of uncultured microbial cells from a sponge sample; 2) isolation of microbial metagenomic DNA; and 3) generation of large DNA fragments of 4 to 10 kb. These resulting large fragments are ready to be cloned into a bacterial host for generating a complex metagenomic library.

indonesian fisheries research journal/indonesian fisheries research journal

ISOLATION AND ENZYMATIC FRAGMENTATION OF GENOMIC DNA FROM UNCULTURED MICROBIAL SYMBIONTS OF Theonella sp. SPONGE AS PREREQUISITE FOR METAGENOMIC LIBRARY CONSTRUCTION