Term
| What are two hormones used in estrus synch? |
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Definition
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Term
| When does oocyte activation occur |
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Definition
| After sperm penetrates zona, before then oocyte was arrested in Metaphase II |
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Term
| You would like to produce a transgenic animal which expresses a certain protein in its milk but not in other tissues. What would you include in the transgene to accomplish this goal? |
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Definition
| tissue specific promoter (casein) |
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Term
If introns are included in a transgene it will not be expressed
T/F |
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Definition
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Term
| When transgenic animals are made by pronuclear injection not all transgenic animals will express the transgene. Why? |
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Definition
| Because random position effects occur Means injection site is random and we don't know what the transgene will express |
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Term
| What is the major advantage of producing transgenic animals by nuclear transfer? |
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Definition
| You can select transgenic cells in culture and all animals should be transgenic |
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Term
| What important transgenic tecnique (or type of experiment), is only possible with embryonic stem cells or nuclear transfer? |
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Definition
| gene targeting (knock out or knock in) |
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Term
| Name two examples of transgenic "bioreactor" systems |
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Definition
homologous recombination
nuclear transfer |
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Term
| Give an example of potential transgenic animal application in agriculture. |
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Definition
| phytase pigs (environmentally friendly) |
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Term
| Give three methods for producing transgenic animals |
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Definition
nuclear transfer
pronuclear injection
embryonic stem cells |
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Term
| What is a tumor inducing (Ti) plasmid? Where does it come from and what is it used for? |
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Definition
| Comes from Agrobacterium tumeficans. Used to produce a tmor in the plant and at the site of injury transforms it to TDNA. At this site you can add the transgene of choice |
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Term
| What is a "Ubiuitous" promoter? Give an example. |
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Definition
| It's expressed in many tissues. CMV, PGK, housekeeping genes |
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Term
| What is siRNA and how is it used? |
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Definition
| Short hairpin RNA, used to regulate expression at the transcription level |
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Term
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Definition
| A gene that has been introduced into the genome of an organism that orginated from another source |
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Term
| Define transgenic organism |
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Definition
| A living organism that harbors an inserted functional gene in its germ line (gametes, passed on through them) |
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Term
| Describe genomic gene structure |
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Definition
| Preferred, not always practical. Exons, plus all introns. Best expression |
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Term
| Describe cDNA gene structure |
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Definition
| Exons only. most available. Low expression gives mixed results, message splicing important. |
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Term
| Describe mini gene structure |
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Definition
| Exons plus one or two introns. mixed results. better than cDNA |
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Term
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Definition
| increase expression levels or tissue specificity. May be an intron. Several kb from promoter |
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Term
| How can you control transgene expression? |
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Definition
High expression in many tissues
Expression in a specific (different) tissue
Gene switches
Promoters and/or enhancers |
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Term
List tissue specific promotors for:
milk
muscle
liver |
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Definition
milk - casein
muscle - actin
liver - metallothioneine |
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Term
| What age are the embryos for nuclear injection? |
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Definition
One cell inject into pronuclei
2-4 cell inject in nuclei |
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Term
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Definition
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Term
| What are methods of producing transgenic animals? |
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Definition
1. nuclear injection
2. somatic cell nuclear transfer
3. embryonic stem cells
4. viral mediated gene transfer
5. sperm mediated gene transfer
6. spermatogonial transfer
7. particle bombardment |
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Term
| How can you identify a transgenic animal? |
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Definition
1. DNA analysis
2. PCR
3. Southern Blot |
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Term
| Problems with nuclear injection |
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Definition
1. Not all animals express the transgene
2. Random positioning effects
3. Establish a line from one or two animals, inbreeding unavoidable. |
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Term
| List the 6 broad classes of GE animals as defined by the FDA |
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Definition
1.To enhance food quality or agronomic traits
2. To improve animal health
3. To produce products intended for human therapeutic use
4. To enrich or enhance the animals interactions with humans
5. To develop animal models for human diseases
6. To produce industrial or consumer products |
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Term
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Definition
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Term
|
Definition
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Term
| What is RNA interference? (RNAi) |
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Definition
| targeting of specific mRNA to inhibit translation |
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Term
| What are the primary steps to the Interference RNA mechanism |
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Definition
1.Long dsRNA is recognized in the cytoplasm by the ribonuclease III-like enzyme DICER.
- DICER cleaves long dsRNA molecules to 21-23 nucleotide dsRNA molecules
- these molecules are known as iRNA
2. iRNA molecules are then incorporated into RISC
- the RISC-iRNA complex recognizes and targets mRNAs for degradation
- effectively stops translation of the targeted mRNA, which is degraded |
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Term
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Definition
RNA
Induced
Silencing
Complex |
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Term
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Definition
small interfering RNA
recently discovered, RISc associate, doesn't require DICER, 21-28 bp long, act as guide sequences for RISC |
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Term
| What is an advantage of using particle bombardment in the production of transgenic plants? |
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Definition
Unlimited host range.
Not limited by ability to regenerate from single cells
Immature embryos from seeds will continue to develop
Transgenic plants selected |
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Term
| What are the 6 genetically modified traits in plants? |
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Definition
- herbicide resistance
- insect resistance
- virus resistance
- altered oil content
- delayed fruit ripening
- pollen control
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Term
| What are two ways transgenic animals could pontentially be important in human medicine? |
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Definition
1. Disease models
2. production of drugs
3. Transplantation of tissue into humans |
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Term
| Describe an example of gene targeting by homologous recombination. |
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Definition
Knock in or Knock out
Mouse embryonic stem cells
Somatic cells |
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Term
Spermatogonial transfer is the most common method of making transgenic animals.
T/F |
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Definition
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Term
| When transgenic animals are made by somatic cell nuclear transfer (cloning), how is the DNA (transgene) usually inserted into cells. |
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Definition
| cloned DNA inserted into somatic or ES cells |
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Term
| How do micro RNA regulate gene expression |
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Definition
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Term
| What is oocyte activation stimulated by? |
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Definition
| fusion of sperm and oocyte plasma membranes causing an increase in free Ca in cytoplasm. |
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Term
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Definition
fusion of parental genomes
pronuclear fusion
replication has occured in pronuclei
immediately into mitosis for divsion to 2 cell |
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Term
| Describe maternal to zygotic transition |
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Definition
transition from using maternal mRNA to zygotic mRNA
8-16 cell bovine
4-8 pig |
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Term
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Definition
Intracellular connections formed (gap jtns)
Outside cells = trophectoderm
Inside cells = ICM
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Term
| Describe Blastocyst Formation |
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Definition
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Term
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Definition
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Term
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Definition
1. Increase # of offspring from selected female
2. Increase # of offspring from selected male
3. Import/Export *Driving force*
4. Treatment of infertility
5. Genectic testing of bulls |
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