Biopanning involves all of the following EXCEPT Screening a series of surface proteins for ability to bind a target such as an antibody. A sequence of 6 histidine at the beginning of each peptide. Use of a phage display library. The bacteriophage M13 All of the above are involved in biopanning.
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Which of the following is correct regarding phage display technology? a. All the answers b. Filamentous phage such as M13 phage is used to replicate in bacteria. c. Eluted phage is usually repassed over the affinity column to isolate the phage with the highest affinity. d. Fully humanized antibodies can be produced using a phage display library. e. In biopanning, the eluting step could be done by the inclusion of a competitive ligand in the buffer.
Adi S.
(h) After selecting a clone carrying the gene for protein P, its cells are propagated to high density in nutrient broth. The chimeric plasmid (with an insert of the gene for protein P) is then extracted and purified from the rest of the cellular DNA. How can we now isolate the gene from the plasmid? (i) How can we demonstrate that the gene we have isolate is indeed the one for protein P? (j) The ampicillin-resistant gene of plasmid pBR322 contains a single PstI restriction site. Genes that are inserted into an amp-r region are highly expressed. If an antibody specific for protein P is available, how would we now construct our gene library and detect the clones of interest? (k) After we have cloned the gene for protein P into the amp^r region of pBR322, suppose that we had to rely only on the drug-resistance properties of the plasmid to select from the gene library those cells carrying the plasmid plus the gene for protein P. What drug-resistance characteristics would the desired cells exhibit?
Sri K.
Researchers have found a human protein (Protein X) that is a useful therapeutic for certain diseases. Protein X doesn't have any significant post-translational modifications and is a relatively small and water-soluble, globular protein. Therefore, Protein X can be expressed in its active form in the bacteria Escherichia coli. Expressing Protein X in and purifying it from E. coli will make it a much more affordable therapeutic. In order to do this, the researchers sequenced the mature mRNA that translates for Protein X in order to get the nucleotide sequence for the mature mRNA that codes for Protein X. They are now going to insert this sequence into a plasmid and transform that plasmid into E. coli, and then express and purify the protein. Which of the following should they also do before they insert the sequence into a plasmid and then into E. coli? Choose one or more: A. If the mRNA contains a UGA stop codon, then change it to a UAA stop codon. B. Add a sequence to encode an affinity tag (such as 6 histidine amino acids) at one end of the gene for Protein X. C. Change most of the AAA codons that encode for the amino acid lysine to AAG codons. D. Synthesize the DNA that is complementary to the mature mRNA sequence, and insert that into the plasmid instead of RNA.
George B.
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