Question #1
only one correct answer per question
part 1-
What was the result of Figure 3?
Only the neo recombinant AAV was replicated in the KB cells with the help of the Ad2 helper virus.
Both recombinant AAV viruses were capable of replication without the help of the Ad2 Adenovirus.
Both the neo and dhfr recombinant AAV were replicated in the KB cells with the help of the Ad2 helper virus.
Neither recombinant AAV virus was replicated.
Part 2-
What was the purpose of constructing a recombinant AAV with the Lambda phage genome?
They wanted to see if they could package the AAV genome into Lambda phage particles
They wanted to see if they could get this bacteriophage to replicate in mammalian cells.
They wanted to use Lambda phage proteins as markers in Western blots
They wanted to construct a recombinant AAV that expressed the capsid proteins that was too large to package.
Part 3-
How did they attempt to produce recombinant AAV virus particles carrying the neo or dhfr genes?
They co-infected human cells with the neo and dhfr AAVs
They infected E. coli with the pBR322 clones.
They co-infected human cells with the neo or dhfr AAVs along with the Lambda AAV clone plus the Ad2 help virus.
Part 4-
How did they titer the wild-type virus used as a control in the recombinant AAV production experiment shown in Figure 4?
They measured AAV genomic DNA by a Southern blot (DNA-DNA hybridization).
They stained cells with an immunofluorescent antibody to detect cells making AAV proteins.
They counted viral particles (virions) by electron microscopy.
Part 5-
What is the result of Figure 4? (see figure above)
Only the dhfr recombinant AAV could be produced.
The recombinant AAV viruses could be packaged when the capsid genes were provided in trans by another AAV virus.
The neo and dhfr recombinant AAVs could not be produced and packaged since they lacked the capsid genes.
Reference
title:
Use of adeno-associated virus as a mammalian DNA cloning vector: Transduction of neomycin resistance into mammalian tissue culture cells
links:
https://europepmc.org/backend/ptpmcrender.fcgi?accid=PMC391945&blobtype=pdf
https://www.pnas.org/doi/pdf/10.1073/pnas.81.20.6466
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