Hello!
I'm a researcher working for identifying novel natural inhibitors for a microbial inhibition. I have selected a target enzyme in the particular species of microbes and now I need to validate whether that target is a suitable one for inhibition.
I am really new to this field of inhibitor discovery and could you please recommend me any methods to do the above step. I have seen there ere many methods of doing it. But most of them seems to be really complex.
The most important thing to do is to make sure that the target is essential for the growth and survival of the microbe. If the target is already well-known, the information may already be in the literature for some microbe, if not the exact one you are interested in.
One way to identify an essential target is to attempt to "knock out" the gene for the target, and to show that it is not possible to obtain any microbes from which the gene has been interrupted. For some microbes, transposon mutagenesis libraries have been created in which an attempt has been made to saturate the genome with transposons to interrupt every gene. The library consists of clones of the microbes that have survived the process. The location in the genome of each clone of a transposon indicates a gene that is not essential. Genes that have not been interrupted by a transposon have a high likelihood of being essential, at least under the growth conditions used. Examining the data from publications on these libraries may be sufficient, if one was made for your microbe.
Article Creation of a Dense Transposon Insertion Library Using Bacte...
Chapter Creating a Library of Random Transposon Mutants in Leptospira
https://www.nature.com/articles/s41576-020-0244-x
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