Hi, I've observed a new biological activity in certain cells, likely induced by an enzyme. How can I determine the gene or genes responsible for expressing this unknown enzyme?
Thanks in advance
One way (probably considered old-fashioned now) would be to isolate the enzyme, obtain a partial amino acid sequence, use the amino acid sequence to prepare degenerate primers, and use the primers to clone the gene from the genomic DNA.
A more up-to-date way would be to obtain the genome sequence of the organism making the enzyme and searching it for genes related to others coding for enzymes with that activity.
Hi Dr. Shapiro,
Thank you for your advice.
One of the colleges tried MS analysis but numerous proteins were present, so it was difficult to identify the sequence.
The genome sequence of the organism making the enzyme is still also not well identified; very few papers about it, I compared the available sequences from yeast with the mammalian genome, and the identity percent was less than 40 %, so I'm not sure if I can depend on this data or not.
Thank you
Alaa
Hello Dear student,
In gene function it can be often obtained by examined it and also where the gene is expressed either in cell or in whole organism.
With determinati of it’s pattern of work and also timing it’s expression can be accomplished by replacing the coding portion of gene under the study with reporter gene.
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