Applications of genetic engineering - lecture 13 Flashcards Preview

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What are the differences between stable and transient transformations?

Transient transfections - short term examinations
stable transfections -allow larger term investigations


How is a transient transfection of a celle done?

- using a lysosomes , with genetic material o the inside
- can use reporter genes to deterine if the genetic material has been taken up such as GFP


What is the significant difference bewtween stable and transient transfections?

stable - DNA incorporated in to the genome
transient - DNA not incorporated into the genome


How can stable transfection occur?

By retrovirus infection


How can you make a retro virus ?

Via a packaging cell- expresses all viral proteins and RNA for the virus to be produced


What is significant about a retrovirus?

Contains reverse transcriptase , so the RNA can be turned into to DNA and incorporated in to the genome


How can you get rid of single genetic mutations?

- Take a slection marker in a new piece of chromosome
- allow the chromosome with the targeted gene and selection marker to go through homologous recomibnation
- selection marker has replaced the mutated sequence


How do you make a chimeric mouse?

- embryonic stem cells from balck mouse , induce modified gene
- These genetically mutated ' black mouse cells' are injected into the blastocyst stage of the embryo from a brown mouse
- Impant embryo into surrogate mother , mother gives birth to a chimeric pup


How can you delete a gene in a specific tissue?

Use the cre- Loxp methodology


How is Haemophilia A caused?

mutations in the gene encoding factor VIII , which is located ono X chromosome


What does Hameophilia A cause?

Allows bleeding to continue as blot clots cannot form


What chromosome is the defected gene on in Haemophilia A?

Chromosome X