New source of cellulase production using a metagenomic technique [electronic resource]

Includes bibliographic reference

The cellulase enzymes with high effectiveness under conditions agreeable to the industrial processes
necessities are one of the keys for the successful development of chemical and drug synthesis. The soil
metagenome is an affluent source for the discovery of new natural products. The objective of the current study
was to identify the isolated functional gene(s) of the cellulase enzyme by using metagenomics. The plan of work
composed of collection of different soil samples, isolation of total DNA, fragmentation, cloning, and expression
of the isolated gene(s) in the suitable host microorganism. The total genomic DNA was extracted using a kit
(QIAGEN), and then digested by different restriction enzymes BamHI. The digested fragments ranging from
~300-5000 bp were ligated, cloned into pUC19 vector, and then transformed into Escherichia coli DH5?. The
resulting clones were screened as cellulase producers using a qualitative method. The positive clones which
showed hydrolysis on the plate were screened once more in Luria-Bertani (LB) medium. The plasmids were
isolated and then tested using universal primer (M13), to detect the fragment size and sequence for the
Polymerase Chain Reaction (PCR) products. This study establishes an effortless and professional method for
cloning of recent cellulase genes through ecological metagenomes. In the outlook, the metagenomic guide
approachs may be functional to the elevated selection of novel cellulase from the environment.

Summary in Arabic