I have a bacterial isolate showing asparaginase activity but till now it is unreported and hence I am unable to find the sequence for asparaginase gene (although, the gene sequence is known in other bacterial genus). I need to clone this gene in E.coli. , what should be my approach to accomplish this?
Cornelia is correct that you can probably design primers just using the most closely related homologs. However if you need a different approach you can get an E. coli strain lacking the native enzyme and set up a select for a clone expressing the enzyme using asparagine for selection.
Thank you all. I have found the protein sequence but its corresponding gene sequence is derived by automated computational analysis using gene prediction method: Protein Homology, can I make the primers based on this gene sequence?
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