Answer bellow remaining questions of Fusion Protein Synthesis and Purification.
ID: 164431 • Letter: A
Question
Answer bellow remaining questions of Fusion Protein Synthesis and Purification.
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Answer only 6 to 11 question
6-What three different techniques did the authors use to lyse these bacterial cells? 7- What is DNase I? Why is it listed in units/ml and not mg/ml? 8-What type of chromatography was used to purify the recombinant proteins? 9-What was used to elute the protein from the beads? 10-What technique did the authors use to concentrate the protein? How did they determine the protein's concentration? 11-Why was propylene glycol added to the protein after concentration?Explanation / Answer
6. Three different procedure for lysis are
7. DNase I, (RNase-free) is an endonuclease that nonspecifically cleaves DNA to release di-, tri- and oligonucleotide products with 5´-phosphorylated and 3´-hydroxylated ends. DNase I acts on single- and double-stranded DNA, chromatin and RNA:DNA hybrids.
One unit is defined as the amount of enzyme which will completely degrade 1 µg of DNA in 10 minutes at 37°C in DNase I Reaction Buffer.
8.Affinity Chromatography by using glutathione sephrose beads to trap GST-tagged protein.
9. Glutathione reduced was used which will work by competetive binding with beads.
10.microcon centrifuge column.
and BSA assay was used to determine the protein concentration.
11. propylene glycol is a cryoprotectant used to preserve the protein for longer time.
11.
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