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1.
a. During development of multicellular organisms the fate of
millions of cells needs to be determined. Transcription factors can
determine cell fate by activating a specific subset of genes available in
the genome. Since transcription factors work either alone as well as in
combination to make such developmental decisions, a large number of
determination events can be specified by a much smaller number of
transcription factors. In theory, how many cell fates could be specified
by four transcription factors?
b. Transcription factors typically recognize a specific binding
sequence (within enhancer elements) to activate or repress a
set of target genes needed to specify cell fate. Describe
different ways that a transcription factor could be involved in
specifying more than one cell fate?
2.
A house fly mutant (ang1-1) lacking antenna was found to have a
mutation in the coding region of a gene encoding a homeodomain
transcription factor. The gene was named ANTENNA GONE1 (ANG1).
a. When and where would you predict the ANG1 gene is
transcribed. What major assumption are you making in your
hypothesis?
b. List at least three methods you could use to test your
hypothesis assuming that ANG1 gene has been identified and
isolated. Which test would you choose to do if you did only
one and explain why?
3.
Three new house fly mutants lacking antennae were discovered
subsequent to the discovery of the ang1-1 mutant. All three new
mutants were mapped to different positions (loci) of the house fly
genome (ANG2, ANG3, ANG4). Two of the genes (ANG2, ANG4) were
identified as encoding new homeodomain transcription factors. It was
found that ANG1 transcript was present in the wild type and ang1-1
mutant but not in ang2 or ang4 mutants. Conversely, ANG2 and ANG4
transcripts were found in each of wild type, ang1-1, ang2 and ang4
mutants.
a. Hypothesize a molecular model that is consistent with the
results described above for the function of ANG1, ANG2 and
ANG4 proteins (include a diagram).
b. A DNA fragment that included the ANG1 promoter and 3000bp
of DNA upstream (away from the coding region) was fused to
the lacZ reporter gene and transformed into flies. Forty
Gene X
A
motif 1
motif 2
motif 3
motif 4
enhancer
promoter exon 1
enhancer
intron
exon 2
5.
The CERX gene of Arabidopsis thaliana encodes a condensing
enzyme required for epidermal wax biosynthesis. The gene has been
cloned (3 exons and 2 introns) and a polyclonal antibody made against
the CERX protein. The antibody was used to show that the CERX
protein is present in epidermal cells but not in other cell types.
a. List three general ways Arabidopsis could achieve such
epidermal specific localization of the CERX protein.
b. Briefly describe how you could use molecular techniques to
differentiate between the three possibilities you listed in part
a.
6.
If cells in a developing C. elegans embryo are ablated with a
laser, typically the adult will lack specific tissues or organs. In
contrast, the same experiment done on an A. thaliana embryo will
normally have little effect on the adult plant. Formulate a hypothesis
that can explain this difference.
7.
You generate a population of enhancer trap lines in the plant
Arabidopsis thaliana by transforming cells with an enhancer trap
construct and regenerating whole plants from single transformed cells.
Each of the transformed plants is heterozygous for a single enhancer
trap inserted in one place in the genome but a different place in each
b.