The operator in an expression plasmid: ______________________ Controls the produ

Question

The operator in an expression plasmid: ______________________

Controls the production of mRNA.

Is the binding location of restriction enzymes.

Is the binding location for RNA polymerase.

Is responsible for directing the mRNA to the ribosome.

Question 2

Select one answer.

10 points

What is the first step that would be required to produce human insulin in bacteria?

Ligate the DNA fragments.

Cut the synthetic gene and expression plasmid with restriction enzymes.

Generate a synthetic gene using chemical synthesis of DNA.

Back-translate the amino acid sequence to a DNA sequence

Determine the amino acid sequence of insulin.

Question 3

Select one answer.

10 points

What is the second step that would be required to produce human insulin in bacteria?

Determine the amino acid sequence of insulin.

Back-translate the amino acid sequence to a DNA sequence

Cut the synthetic gene and expression plasmid with restriction enzymes.

Generate a synthetic gene using chemical synthesis of DNA.

Ligate the DNA fragments.

Question 4

Select one answer.

10 points

What is the third step that would be required to produce human insulin in bacteria?

Determine the amino acid sequence of insulin.

Generate a synthetic gene using chemical synthesis of DNA.

Back-translate the amino acid sequence to a DNA sequence

Cut the synthetic gene and expression plasmid with restriction enzymes.

Ligate the DNA fragments.

Question 5

Select one answer.

10 points

What is the fourth step that would be required to produce human insulin in bacteria?

Determine the amino acid sequence of insulin.

Cut the synthetic gene and expression plasmid with restriction enzymes.

Generate a synthetic gene using chemical synthesis of DNA.

Back-translate the amino acid sequence to a DNA sequence

Ligate the DNA fragments.

Question 6

Select one answer.

10 points

What is the fifth step that would be required to produce human insulin in bacteria?

Cut the synthetic gene and expression plasmid with restriction enzymes.

Generate a synthetic gene using chemical synthesis of DNA.

Ligate the DNA fragments.

Determine the amino acid sequence of insulin.

Back-translate the amino acid sequence to a DNA sequence

Question 7

Select one answer.

10 points

Which of the following recognition sequences will produce sticky ends (the cleavage site is marked by ^).

^AAATTT

AAATT^T

AA^ATTT

All of the other selections.

Question 8

Select all that apply.

10 points

The DNA sequence of the expression plasmid adjacent to the ribosome binding site is:

---GGATCCATCTGCAGATCT----

---CCTAGGTAGACGTCTAGA----

It contains a BamH1 site (G^GATCC) and a BglII site (A^GATCT). In order for the synthetic gene to be inserted between these two sites, it should be constructed to have which restriction sites at its beginning and at its end? Select all that apply.

EcoR1 (G^AATTC), followed by BamH1 (G^GATCC).

BamH1 (G^AATTC), followed by EcoR1 (G^AATTC).

BamH1 (G^GATCC), followed by BamH1 (G^GATCC).

BamH1 (G^GATCC), followed by BglII (A^GATCT).

BglII (A^GATCT), followed by BglII (A^GATCT).

Question 9

Select one answer.

10 points

Examine the three base universal codon table below to answer this question.

The beginning part of the sequence of human insulin is Met-Ala-Leu-Trp. Which of the following sequences would be ideal for the production of human insulin in yeast?

ATG GCG CTG TGG

ATG GCC CTT TGG

ATG GCT TTA TGG

Question 10

Select one answer.

10 points

The following is the DNA sequence of the expression plasmid adjacent to the ribosome binding site:

-GAATTCGATATCAAGCTT

(EcoR1)(EcoRV) (HindIII)

You generated a synthetic gene for a human protein, but unfortunately you forgot to include the recognition sequences for EcoR1 at the beginning and for HindIII at the end of your synthetic gene, thus your gene looks like:

AGT------TAA

TCA------ATT

Given the following recognition sites: EcoR1 G^AATTC, EcoRV GAT^ATC, Hind III A^AGCTT, can you still insert your synthetic gene into the expression plasmid?

No

Yes

Explanation / Answer

1)

The operator in an expression plasmid Controls the production of mRNA by allowing the biding of repressor proteins. Thus, the correct option is (a).

2)

The first step that would be required to produce human insulin in bacteria is cut the synthetic gene and expression plasmid with restriction enzymes. Thus, the correct option is (b).

3)

The second step that would be required to produce human insulin in bacteria is Ligate the DNA fragments. Thus, the correct option is (d).

4)

The third step that would be required to produce human insulin in bacteria is Generate a synthetic gene using chemical synthesis of DNA. Thus, the correct option is (b).

5)

The fourth step that would be required to produce human insulin in bacteria is Determine the amino acid sequence of insulin. Thus, the correct option is (a).

6)

The fifth step that would be required to produce human insulin in bacteria is Back-translate the amino acid sequence to a DNA sequence. Thus, the correct option is (e).

7)

AAATT^T, this is the proper palindromic sequence can generate sticky ends upon restriction enzyme digestion.   

8)

BamH1 (G^GATCC), followed by BglII (A^GATCT), then only the new gene properly inserted into the plasmid.

9)

ATG GCG CTG TGG, this is the proper DNA sequence of these amino acids.

10)

No, this is because the sticky ends are formed by the digestion only EcoRV site.    

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