Your assignment is to describe in detail how you would make a transgenic animal

Question

Your assignment is to describe in detail how you would make a transgenic animal that expresses the human gene for the novel Protein KTH (does not actually exist) specifically in its milk. Protein KTH is functional as a glycosylated hetero.dimeric protein.

a) You may choose the species in which you will express the gene (E. coli, mouse, cow, pig, horse, sheep, goat, etc.). Explain your choice in detail.

b) You should provide an illustration of the genetic elements that you would use, in the order in which you would use them( include a detaileg explanatin of the why you used each element; provide refrences for each sequence).

c) Describe in detail how you will determin which offspring are transgenic and appriximately how many copies of the gene are present in the offspring.

d) Congradulations! Your animals are positive for the transgene! what steps do yoy need to take now before you breed a whole bunch of animals? Hint: how would determine if the protein is expressed only where trageted?

e) Is an animal model or animal cell model required to express this particular prottein ? Can it be properly expressed in E. coli? Explain in detail.

THIS IS A GENETIC COURSE QUESTION!

Explanation / Answer

a. Suitable organism = Pig or sheep. They are eukaryotic animals that can produce large amounts of milk. The cow also can be used but it has larger generation time.
b. The genetic elements to be used = Highly active promoter, an element that provides tissue-specific expression, coding sequence, and tag. Other general elements such as transcriptional start and stop site must be included in the construct.
c. We can perform PCR with transgene-specific primers to determine whether they are positive for the transgene. We can perform Southern blot to determine the copy number.
d. We should isolate protein samples from each tissue and perform western blot with specific antibody to detect in which tissues the protein is being expressed.
e. Since the active protein requires post-translational modifications, it can not be expressed in E coli.

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