Please help with this biology question: You have discovered a human chemokine re
ID: 9125 • Letter: P
Question
Please help with this biology question:You have discovered a human chemokine receptor and have identified a 500bp sequence. You have decided to insert this sequence into the PGEM-T Easy vector, a 3200bp plasmid. You produce a PCR product corresponding to the above 500bp sequence, and using electrophoresis, confirm the presence of a single 500 bp band. You next set up a ligation reaction using this PCR product and PGEM-T Easy, and transform competent JM109 bacterial cells with your ligation product. You then select for transformed cells.
Q: For your experiment you must have a plasmid with a single copy of the PCR product inserted into the PGEM-Easy vector. How would you CONFIRM that you have selected cells carrying the recombinant plasmids with this specific configuration?
Explanation / Answer
howdy fun question - you have to think like a real geneticist and microbiologist to get these. im not sure if this is what your teacher is looking for, but very often when performing htese kinds of experiments, another gene that encodes antibiotic resistance to a particular drug is included in the plasmid. that way, you know a bacterial colony has succesfully transformed the plasmid if it is able to grow in the presence of that particular antibiotic. any bug that didnt pick up the resistance gene along with the plasmid of interest would have died. hope that helps! jonathan