2.) if you wanted to express a large eukaryotic gene in a prokaryotic organism, what steps would be necessary? describe your protocol below including the suitability of a vector for a large gene.

Explanation:

The eukaryotic genes of interest can be expressed in the prokaryotic genome or plasmid vector. The following are the steps used for the purpose of expression of eukaryotic genes in the prokaryotic genome.

1. Amplification of the target gene of interest from the eukaryotic genome by PCR or polymerase chain reaction.

2. The selection of the plasmid and digestion of the plasmid DNA with the restriction enzyme.

3. Ligation of the amplified DNA fragment and the plasmid.

4. Introduction of the ligated plasmid into the prokaryotic host organism.

you are certainly talking about this picture:

a: is the label of the white square, it is showing the  replication fork, formed by helicase (d) ==> so it is the site of dna separation and replication 

b: is a new dna brand that is following directly the replication fork, so it is continuous ==> built continuously from 5' to 3

c: dna polymerase cannot replicate dna on its own, it needs the of their proteins, and one of them plays the role of a clamp which fixes  the dna with the dna template and increases  the processivity of the enzyme. ==>   stops polymerase from falling off the  template.

d: is the enzyme responsible for forming the replication fork (helicase) ==> unwinds and separates dna into two strands

e: is the delayed strand, it is built  in the opposite sense of the helicase, so dna will be built in fragments ==> built  fragments from 5' to 3 joins

and finally f: is the enzyme that builds  the new dna strand (dna polymerase) ==>   adds nucleotides to make a new dna helix.

credit: gradman555p98x9p

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