Please answer all of them. Only 3-4 sentences. the strategy for the expression o
ID: 267910 • Letter: P
Question
Please answer all of them. Only 3-4 sentences.
the strategy for the expression of a target protein in a host organism involves 11. Sometimes synthesizing the protein as part of a fusion protein. Why is this approach useful? How is a fusion protein created? 12. How would you manipulate the DNA sequences upstream of a target gene to modulate expression of that gene? 13. How would you avoid some of the problems associated with the limited amount of oxygen that is available to growing E. coli cells when a foreign protein is overproduced?Explanation / Answer
11. fusion protein is a recombinant protein formed by joining two three genes coding for other different proteins. this approach helps in stable transformation of target gene in recombinant cells. it is created by removing stop codon from cDNA end of first gene which is then ligated with second gene and so on. it is also used as tags in cell biology example GFP tags.
12. upstream sequences can be modulated as efficient promoters which promotes rapid transcription of target genes they can be enhanced by adding certain factors or genetic sequences to promote high transcription rates.
13. additional oxygen can be provided and stirring can be done to properly mix the culture with nutrient and gases also, by removing overly produced proteins can enhance the diffusion rate of oxygen into the culture
18. different subunits can be packaged and produced in different e coli cells and later can be combined to form an active protein product one such example is production of active insulin from E coli cells in which both alpha and beta subunits are produced separately then combined with active disulphide bridges in vitro.
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