Question

For a project in my microbiology class, I was asked to come up with a scientific question about GP187 (Singulisphaera) and the gene for MoxR proteins. I came up with the following: "I discovered that MoxR is the key regulator of the protein methanol dehydrogenase, which is important in the utilization of methane and methanol. How does the addition of methanol to the MoxR protein gene affect the function of the gene and the production of methanol dehydrogenase in GP187 (Singulisphaera)?" However, I am supposed to create a hypothetical research plan to answer this question and I do not know how to tackle that. How would I do this in terms of specific laboratory procedures and techniques? I basically need to propose a research plan to answer the question and I have no idea how to do so. The last person that answered this said that it was out of their expertise. Please help if you can

Answer 1

If MoxR is affecting the expression of the gene of methanol dehydrogenase, you need to check the production of methanol dehydrogenase in MoxR+ cells and MoxR- cells in presence of methanol in the medium. From this experiment you would know that MoxR is positive regulator or negative regulator (if it is positive regulator : methanol dehydrogenase(MD) will not get produced in MoxR- cells, and if negative : MD will get produced in both type of cells.

You can create MoxR- cells by various techniques such as site directed mutagenesis, transposon mediated mutagenesis, CRISPR-CAS9 etc.