The assignment is due on December 8. It is to be submitted by email before the end of the class lecture period
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Description
Due date: The assignment is due on December 8. It is to be submitted by email before the end of the class lecture period (i.e., by 5:45 pm on December 8). Late assignments will be penalized according to the guidelines found in the course outline. Your assignment must reflect an independent effort.
Assignment: You have recently graduated from university and got a job with a small upstart
biotechnology company. You have been hired because of your expertise in molecular and cell
biology. You have been assigned to the SARS-CoV-2 vaccine project. Your boss wants to copy
the Moderna approach and target the 1273 amino acid, spike glycoprotein (coded by gene S) as
the vaccine antigen.
He wants you to make two different expression constructs and confirm the products
from these expression constructs. Expression construct 1 will produce the spike glycoprotein
with an N-terminal 6xHis tag. In contrast, expression construct 2 will result in the production of
spike glycoprotein mRNA. You only have access to pET28b (See map on Moodle site or in
appendix of this assignment) for making the constructs. Your boss also wants you to use only
Escherichia coli as the expression host. You can use PCR but not DNA synthesis (your project
does not have the budget for this). You have been given about 1 g of genomic ssRNA (positive
sense) from novel coronavirus SARS-CoV-2 isolate Wuhan-Hu-1 (GenBank Accesssion number
NC_045512). You have access to a well equipped molecular biology laboratory with a modest
budget. You can have primers synthesized in house. There is also an in-house group that will do
automated Sanger sequencing as well as next gen sequencing with a Nanopore PromethION
instrument.
