The researchers from the previous method made several constructs of ARD1 to stud
ID: 953406 • Letter: T
Question
The researchers from the previous method made several constructs of ARD1 to study the various domains, which are shown below, and numbered 1-6. Match the construct (1 through 6) with the appropriate lane on the SDS-PAGE gel to the right. Enter the numbers above each lane. Each protein shown is fused to GST (26 kDa, not shown). An amino acid’s Mw, on average, is 112 Da. The numbers on top of the constructs (i.e. 1, 402, 574) indicate the amino acid number (which should allow you to calculate the approximate Mw).
31. _______ _________ _________ _________ _________ _________
32. Some lanes show a contaminant. What is the contaminant at the bottom of lanes 1 through 5? __________
What event likely produced these products? __________________________________________________
Explanation / Answer
In SDS PAGE due to SDS protein denatures and only molecular weight directs the separation.
Average amino acid weight is 112 Da.
1.ARF – 403 to 574 has 171 amino acids. Its molecular weight is 171*112= 19152 Da = 19.1 kDa
2. 88 to 574 has 486 amino acids. Its molecular weight is 486*112=54432 Da = 54.4 kDa
3. 88 to 402 has 314 amino acids. Its molecular weight is 314*112 = 35.16 kDa
4. 1 to 110 has 110 amino acids. Its molecular weight is 110*112 = 12.32 kDa
5. 1 to 402 has 402 amino acids. Its molecular weight is 402*112 = 45.02 kDa
6. 1 to 574 has 574 amino acids. Its molecular weight is 574*112 = 64.28 kDa
Numbers on lane are
6 – 2 – 5 – 3 – 1 – 4
32.
Contaminant on some lanes is GST. Which is commonly used for tagging.
Special elusion methods should be used for GST elusion.