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Unformatted text preview: f the knockout mutation. Which of the
following steps will you do and in what order?
i. Transform haploid a cells.
ii. Transform haploid cells.
iii. Select KanR cells.
iv. Transform diploid a / a cells.
v. Transform diploid a / a cells.
vi. Sporulate the transformed cells.
vii. Mate the transformed cells with cells of the
opposite mating type.
viii. Add the KANR gene to the plasmid vector.
ix. Clone the KANR gene into the H2A gene on
b. You have obtained haploid cells that have the H2A
gene knockout but there is no phenotype. The cells live and appear to grow normally. Offer two possible explanations for this result.
c. How could you determine if there is more than one
copy of the H2A gene in the yeast genome?
d. You located a second copy of the H2A gene, determined the sequence, and found that the two copies
had only a few base differences. Transcription of
the ﬁrst copy of H2A that you isolated is transcriptionally regulated during the cell cycle. How could
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- Spring '08