Sequencing
Transgenes
Knockout/gene replacement
Transgenic Animals
100
What type of sequencing utilizes overlap of small fragments to compile the genome sequence?
Shotgun sequencing
100
What does transgenic mean?
Adding foreign DNA to the genome
100
What is gene replacement?
correcting a mutant strain
100
What are three reasons to make a transgenic animal?
to look at expression pattern of a gene, label a particular cell, overexpress a normal or mutant gene
200
This type of sequencing uses newly synthesized fluorescently labeled nucleotides to determine the order one base at a time.
Next gen sequencing
200
The genetic code is ______ between species, and the promotors and regulators are ______ between species.
the same; different
200
What is the purpose of gene knockout?
to determine the function of a gene by disrupting it and analyzing the resultant phenotype
200
What is GFP used for?
green fluorescence protein; used for labeling
300
How does comparing cDNA with the entire genome allow you to tell where the introns and exons are?
cDNA is made from reverse transcribed RNA, so it has no introns
300
Name two factors that can negatively affect heterologous protein synthesis.
codon bias, differences in post-translational modifications, aggregation/precipitation of inactive protein
300
Of the four meiotic products, how many would contain the disrupted gene?
two
300
How do you insert the transgene into the animal's genome?
in vitro fertilization; micro-inject DNA into the pronucleus of a fertilized egg, then introduce the fertilized egg into a foster mother
400
What does a genomic library have that a cDNA library lacks?
promotors, enhancers, introns, repetitive sequences
400
To make transgenic yeast using a single crossover, the ___ is is not integrated with the plasmid.
origin of replication
400
If the spores with disrupted genes failed to grow when incubated, you could assume the gene is ____.
essential
400
What are two disadvantages resulting from random integration sites of foreign DNA in transgenic mice?
can change the expression of your gene of interest based on where it integrates; can create unrelated mutant phenotypes by disrupting other genes
500
Why would genomic libraries taken from two different tissues of an organism be the same, but cDNA libraries taken from two different tissues be different?
genomic libraries contain all genes, cDNA libraries contain only the expressed genes, which would differ by tissue or stage of development
500
What does a terminal deletion mean in reference to a single crossover transformation?
Host loses all of its chromosome distal to the integration site
500
What is a potential problem that could arise by using a single crossover to make a knockout?
You would eliminate parts of the chromosome distal to your integration site, which could contain essential genes, so you couldn't be sure if the gene you were studying was essential or if it was another one downstream
500
Expression of transgene is often _____ than normal because___?
higher; multiple copies of the transgene are integrated and expression is uncontrolled
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