sure your answers are hand written. Keep your responses reasonable... usually a
ID: 210499 • Letter: S
Question
sure your answers are hand written. Keep your responses reasonable... usually a few sentences will get the iob done. 1. OK. So, let's go back to CandyP, the delicious eukaryotic protein we talked about in class that tastes like a Snickers bar. a) You painstakingly derived the (mature) transcribed sequence for CandyP, part of which is listed below. What would be the mRNA sequence that corresponds with this segment? 2 template 5' TACCGTACGTAAC3 I coding3' A TGGCATGCATT G5 3 OK, how you need to get E. coli to make a whole bunch of that protein. What do you need to do to this DNA sequence before inserting it into a plasmid for transformation, to make sure that the E. coli will make this protein? Why? b)Explanation / Answer
a. The mRNA sequence is same as that of coding strand of DNA. Only difference is that in place of thymine uracil is present. Hence mRNA sequence will be:
5' AUGGCAUCGAUUC
b. E. Coli can not take external DNA as it can not pass through cell membrane. Firstly, E. Coli cell are made competent with the help of specific concentration of divalent cations viz calcium chloride and magnesium chloride. After that, E. coli cells are incubated with recombinant DNA on ice, followed by giving the cell heat shock treatment and then again place it in a DNA.
We need to do for expression of the the protein
1. Expression plamid vector for expression of protein
2. Rare tRNA should be incorporated to overcome codon bias.
3. Chaperon should be included in order to fold the inserted protein.
4. Solubility enhancing fusion tags should be incorporated
2.
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