Biol 2104: Lecture 20- Methods In Molecular Biology

Front 1

What are expression vectors, and what are they used for?

Back 1

expression vectors are vectors with a highly active promoter, causes large amounts of mRNA to be produced when a protein-coding gene is inserted

Used for: production of large amounts of protein

Figure 10-32 (353)

Hint 1

protein

Front 2

What are reporter genes?
What are they used for?

Back 2

reporter genes are genes that have a coding-sequence for a reporter protein (such as GFP).

Used for: tracking the level, timing, and cell specificity of expression of reporter protein production. This gives clues about the function of the original gene (replaced by the reporter gene) and the action of its regulatory DNA sequences.

Figure 10-34 (354)

Hint 2

regulatory DNA sequences

Front 3

Green fluorescent protein (GFP)

Back 3

A commonly used reporter protein.

Figure 10-35 (355)

Front 4

site-directed mutagenesis

Back 4

technique by which a mutation can be made at a particular site in DNA (changing AA sequence--> causes various changes in the resulting protein)

-By changing selected AAs in this way, on can determine which parts of the polypeptide chain are crucial to various properties of a protein (such as folding, protein-ligand interactions, catalysis- in the case of enzymes).
-Allows one to deterimne the biological roles of each part of a given protein

Figure 10-36 (355-6)

Front 5

transgenic organism

Back 5

and organism into which a new gene has been introduced, or whose genome has been altered in other ways using recombinant DNA techniques

356-7

Front 6

3 types of gene alterations that can be used to alter the genome of an organism (-->transgenic organism)

For each case, which gene(s) will be active?

Back 6

1) Gene Replacement
2) Gene Knockout
3) Gene Addition

1) only mutant gene active
2) no active gene present
3) both genes active

Figure 10-37, also read (don't have to memorize all of it tho) chart drawn in notes