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A. Right
but wrong -
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A. Right
but wrong -
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All vectors must contain
an origin of replication, so this characteristic cannot serve as a basis
for vector selection.
Try
again
B. Right
- this will allow you to adopt a one step purification procedure. You will
still have to decide whether you use a vector that allows the expression
of the His tag at the N-terminus or at the C-terminus of the protein*.
*
Close proximity of the His-tag to the active site might
alter essential characteristics of the fusion protein. Hence, the His-tag
should be fused, where it stands the smallest chance to interfere with
the protein's activity, the protein's folding, and the fusion protein's
capacity to bind to the nickel column during purification.
C. Wrong - RNA polymerase is present in the host cells. It is not necessary to include it in the vector. Sometimes host cells are specially engineered to contain a specific RNA-polymerase, that is not normally present in them to increase the specificity of the induction procedure.
D. Wrong-
since we suspect that BHR1 protein over-expression may be toxic to the
host, it is better to induce its expression as late as possible, in order
to allow virus formation. Therefore, it is better to use a very late promoter.
Moreover,
ALL baculovirus vectors are based on late promoters, to allow large
scale production of proteins, and so are suitable for the production of
toxic proteins.
E. Right - since you wish to create a His tag fusion protein, it is essential to place the BHR1 sequence and the poly-His encoding sequence in frame.
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