BIO130H1 Lecture Notes - Lecture 11: Reverse Transcription Polymerase Chain Reaction, Reverse Transcriptase, Cleavage And Polyadenylation Specificity Factor

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Producing many proteins from one rna sequence due to the introns. If you are careful where you design your primers, you can analyze which exons are included in the mrna. Studying which genes are transcribed and how much are. After the rna polymerase has become proccessive, the 5" capping protein hopp on the mrna and put on the 5" cap. Eventually, the rna polymerase will hit a consensus sequence which is a termination sequence. It doesn"t recognize the termination signal sequence and thus keeps transcribing and adding on nucleotides. The 3" processing protein (that have been attracted to the tail) will recognize the termination sequence. This is when the cleavage of the sequence will occur. Transcription of the consensus sequences (termination sequence) & recruitment of modifying. Proteins: phosphatases in the cell will dephosphorylate the dna template and thus the rna polymerase will then fall off after the termination sequence (consensus sequence)

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QUESTION 1

Which of the following statements regarding splicing is FALSE?

A) The length of introns determines the efficiency of splicing.

B) Many human neurological disorders are caused by splicing errors and/or mutations in splicing factors.

C) Splicing requires the action of a variety of snRNAs that direct the transesterification reactions.

D) Splicing is dictated by sequence features in pre-mRNA transcripts

QUESTION 2

Which of the following mRNA processing factors does NOT associate with the CTD of RNA polymerase II?

A) splicing factors

B) polyadenylation factors

C) miRNAs/siRNAs

D) capping proteins

QUESTION 3

Which of the following statements about mRNA stability is FALSE?

A) Deadenylation-independent mRNA decay is exclusively in the 5' to 3' direction.

B) The most common pathway of mRNA decay is deadenylation-dependent.

C) Yeast mRNA decay occurs primarily in the 5' to 3' direction, while 3' to 5' decay is more common in mammalian cells.

D) Decapping enzymes are required for all pathways of mRNA decay.

QUESTION 4

You obtain the sequence of a gene containing 10 exons, 9 introns, and a 3â² UTR containing a polyadenylation consensus sequence. The fifth exon also contains a polyadenylation site. To test whether both polyadenylation sites are used, you isolate mRNA and find a longer transcript from muscle tissue and a shorter transcript from all other tissues. Which of the following is the most likely mechanism underlying these alternative transcripts?

A) The fifth exon is spliced out of the mature mRNA transcript in muscle cells.

B) Muscle cells have less effective polyadenylation than other tissues.

C) An alternative CPSF protein is expressed in muscle cells with lower affinity for the polyadenylation sequence in exon 5.

D) The mRNA in muscle cells is edited to eliminate the polyadenylation sequence in exon 5.

blushdonkey474

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

1)What enzyme is used to attach different colors of stained nucleotides to create a DNA molecule along the mRNA?

Select one:

a. reverse transcriptase

b. fluorescent primer

c. restriction enzyme

d. radioactive probe

The process in which two complementary strands of DNA from different sources pair with one another is called hybridization.

Select one:

True

False

3) Explain the significance of a red microarray spot revealed by the microarray scanner.

Select one:

a. this gene is not expressed in either type of cell

b. this gene is expressed in cancer cells

c. this gene is expressed in healthy cells and cancer cells

d. this gene is expressed in healthy cells

4)What is the function of primers in PCR?

Select one:

a. raw material for new DNA

b. attach new nucleotides to DNA strand

c. attach to either side of target DNA

d. denature, or separate, DNA

5)What is the function of DNA polymerase in PCR?

Select one:

a. raw material for new DNA

b. attaches to either side of target DNA

c. attaches new nucleotides to DNA strand

d. denatures, or separates, DNA

BIO130H1 Lecture Notes - Lecture 11: Reverse Transcription Polymerase Chain Reaction, Reverse Transcriptase, Cleavage And Polyadenylation Specificity Factor

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