Question

Through genetic engineering, Escherichia coli becomes capable of biofuel production by consuming sugars derived from plant biomass. However, E. coli suffers from low tolerance to growth-inhibitory compounds in the culture medium, such as hydroquinone, which is generated during the pre-treatment of plant biomass. Therefore, you will introduce new genes to E. coli, in order to increase its tolerance to hydroquinone and thereby improve the biofuel production. To identify these genes for better tolerance to hydroquinone, you want to screen DNA molecules present in environmental soil samples. Describe each step toward identifying gene sequences that will give higher tolerance of E. coli to hydroquinone.

Answer 1

Functional metagenomics analysis can be used to identify the gene sequences that will give higher tolerance to hydroquinone.

For that, First isolate the DNA from the soil sample. Create a library of these isolated sequences by cloning these extracted DNA sequences into a suitable vector. Then transform Ecoli cells with the clones in the library. Screen and select those transformed colonies by culturing the cloned Ecoli in media containing hydroquinone. Plasmid containing the gene can be extracted from the positively transformed colonies, so that the gene can be isolated by PCR amplification and sequenced.