BIOL 2002 Chapter Notes - Chapter 20: Chain Reaction, Polymerase Chain Reaction, Reverse Transcriptase

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indigoraccoon472

2 Apr 2017

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Make many copies of a gene to study it directly or to use it as a tool. Study gene sequence to see if it has a mutation in a diseased person. Want a lot of the gene product. Helpful for molecular biology, genetics, cell biology, biochemistry, medicine. Isolate vector dna from bacterial cell and gene of interest from chromosomal dna. Introduce a recombinant vector into a host cell that does not already have a vector. 3: the host cell takes up onie vector and then copies the vector and divides to produce many cells. Vector dna acts as a carrier of the dna segment to be cloned. A vector can carry a small segment of the dna or a single gene. When a vector is introduced into a living cell, it can replicate so the dna it carries can also be replicated. Plasmids - small, circular pieces of dna found in bacteria.

Related Questions

1. You have cloned a human gene along with its promoter and ribosome-binding site in a bacterial cloning vector and then introduced the recombinant plasmid into a bacterial host cell that supports the replication and segregation of the cloning vector. Unfortunately, even though you know the whole human gene sequence is present (by DNA hybridization); you cannot get expression of the gene. Which of the following is NOT a reason why there is no expression?

A. The vector cannot replicate

B. The human promoter is not recognized by the bacterial host cell

C. The ribosome-binding site is not recognized by the bacterial host cell

D. The human gene lacks introns, so it is not transcribed by the host bacterium

2. Restriction endonucleases are:

A. sequence-specific

B. able to recognize any 4-, 6-, or 8-base pair sequence

C. always able to produce single stranded tails

D. only able to cleave bacterial DNA

3. Which of the following is NOT a required characteristic of cloning vectors?

A. Genes for replication of vector DNA and segregation to progeny cells

B. Genes for selection of cells carrying the vector (e.g., antibiotic-resistance)

C. DNA sequence for the introduction of foreign DNA where the integrated DNA does not disable the vector

D. None of the above

4. A poly (T) primer is used to produce cDNA from mammalian mRNA. Which of the following would be used to produce cDNA from bacterial mRNA?

A. A poly (T) primer

B. A random DNA sequence primer

C. A poly (A) primer

D. A poly (GC) primer

Recombinant DNA I: Core Question 1a (1 point possible)

Why is biochemical purification of one gene away from the othergenes in a cell more challenging than biochemical purification ofone protein away from the other proteins in a cell? -unanswered

a)A cell has many more genes that it has proteins.

b) DNA is very unstable and is likely to be broken down duringthe purification process.

c) Proteins are larger molecules than genes.

d)The chemical nature of all genes is very similar; the chemicalnature of various proteins can be very different.

DNA I: Core Question 1b (1 point possible)

Which of the following are true of a vector, as described in thelecture? Check all that apply. -

- A vector is made of DNA

-A vector can be replicated by the host cell

-A vector can be put into a host cell by transformation

-A single copy of the vector can be found in each transformedcell

orchidrat219

Recombinant DNA Technology

The questions all relate to the same topic and somewhat interrelate

1. Plasmids used in vitro to clone foreign DNA fragments are called ___

A. Transgenic.

B. Clones.

C. Vectors.

D. Conjugants.

2. VNTR alleles are hypervariable regions of human DNA that differ from each other in:

A. Location of internal sites recognized by restriction enzymes.

B. Variable number of point mutations.

C. Number of copies of an internally repeated DNA sequence.

D. Variable location on different chromosomes.

3. Probability calculations are used in forensic applications of DNA fingerprinting to determine if:

A. DNA from two different sources has matching alleles.

B. DNA samples were degraded before analysis.

C. A match between alleles of different DNA samples might have occurred by chance.

D. An unknown DNA sample is from a male or a female.

4. Plasmid vectors used in cloning often contain a gene for the N-terminal 146 amino acids of the enzyme Î²-galactosidase. What is the purpose of including this gene in the vector?

A. It allows selection of E. coli host cells that contain the plasmid.

B. It allows selection of E. coli host cells that contain plasmid in which the insert has been ligated.

C. It cleaves the insert to allow it to be ligated into the vector.

D. It enables the plasmid vector to replicate in E. coli host cells.

5.The first gene therapy clinical trial took place in 1990, on patients with SCID (severe combined immunodeficiency syndrome). Is this true or false?

A. True

B. False

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BIOL 2002 Chapter Notes - Chapter 20: Chain Reaction, Polymerase Chain Reaction, Reverse Transcriptase

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