Which Technology is valuable for mass producing drugs and other useful proteins?

Recombinant DNA Technology

What type of probe would you use for a western blot experiment?

An antibody

Which matrix can be used for protein gel electrophoresis?

Polyacrylamide

What technique is used to look at the expression of thousands of genes simultaneously in 1 experiment?

DNA Microarrays

Approximately how large is the human genome?

3.1 billion bp

What is the study of the entire set of proteins expressed by a genome, cell, tissue, or organism?

Proteomics

Austrian monk who worked with inheritance of traits in peas and is considered the father of genetics.

Gregor Mendel

The entire genetic code consists of how many codons?

64

What does it mean that the genetic code is redundant?

2 or more triplet codons on an mRNA can code for the same amino acid.

What is true of RNA polymerase?

Moves along the template DNA strand in a 3' to 5' direction.

What is a codon?

3 nucleotide sequence that codes for a specific amino acid.

What is BRCA1?

A defective gene in breast cancer.

What is the role of a spliceosome?

Removes introns from a transcribed pre-mRNA.

What was the first recombinant drug approved by the FDA?

human insulin (Humulin)

Who discovered Penicillin?

Alexander Fleming in 1928

Agricultural/plant biotechnology

Green Biotechnology

Medical/pharmaceutical biotechnology

Red Biotechnology

White biotechnology

Industrial biotechnology

Nano Biotechnology and bioinformatics

Blue Biotechnology

In DNA, which base pairs have 2 H bonds?

A-T

The set of all RNA molecules, including mRNA, rRNA, tRNA, and non-coding RNA produced in 1 or a population of cells

Transcriptome

Who was the Scottish embryologist that created the first true clone, the Dorset ewe Dolly in 2003?

Ian Wilmut

The first synthetic cell created by transferring the whole synthetic genome into M. capricolum cell was made in 2010 by an American scientist _______.

J. Craig Venter

Name structures of a plant cell that are not in an animal cell.

Chloroplasts, Cell Wall, large central vacuole (as opposed to many small ones in animal cells)

What is Southern blotting?

DNA analysis that allows for the detection of a specific DNA fragment in a complex mixture.

What is Western blotting?

Protein analysis for separating protein molecules by gel electrophoresis and transferring proteins onto a filter paper blot that is usually probed with antibodies to study protein structure and function.

What is Northern blotting?

RNA analysis for separating RNA molecules by gel electrophoresis and transferring RNA onto a filter papaer blot for use in hybridization studies.

Which enzymes recognition sites would produce sticky ends compatible with (5'-GIGATCC-3')

BamH 1 (3'-CCTAGIG-5')

What effect would you expect if gene expression of lac operon were completely repressed?

Lactose would not be converted into inducer and operon could not be produced

What is RT-PCR (Reverse transcriptase-polymerase chain reaction)?

Technique for studying gene expression (not actual genome).

Uses the enzyme reverse transcriptase to copy RNA from a cell into cDNA and then amplifying cDNA by the polymerase chain reaction.

What is qRT-RT-PCR (real time or quantitative PCR)? How is it different than RT-PCR?

Modern PCR technique that uses dyes to measure the amount of PCR product made as it is occurs.

Different than RT-PCR b/c results are obtained throughout the experiment rather than after.

What is the most commonly used stop codon?

UGA

Best expression host for producing candidate therapeutic human proteins that require post-translational modification for recombinant proteins to be active and functional will be...

Mammalian cells

_____ encoded enzyme breaks down ampicillin that allows for easy selection of bacteria on plates that took up plasmid after transformation.

Antibiotic resistance gene ampR

T/F: For transcription RNA polymerase does not need a primer.

True

In 1968, who discovered plasmid?

Stanley Cohen in 1968

Which human chromosome contains over 4,000 genes?

Chromosome 1

T/F: Taq polymerase puts a single adenine nucleotide on 3' end of all PCR products.

True

Explain ELISA

Stands for Enzyme-Linked Immunosorbent Assay. It uses antibodies and immunoassays to detect specific antigens in a sample.

A complex of DNA and proteins that forms chromosomes within the nucleus of eukaryotic cells.

Chromatin

Primary Protein structure

The sequence of a chain of amino acids/peptide bonds.

Secondary protein structure

The sequence of amino acids is linked by weak Hydrogen bonds into either a Beta sheet structure or an Alpha Helix structure.

Tertiary protein structure

Begins when certain attractions and interactions occur between Alpha helixes and Beta sheets.

Quaternary protein structure

Some proteins consist of more than 1 amino acid chain, each participating in the final 3D shape.

Which organelle is the site of synthesis of secretory proteins from bound ribosomes, and where carbohydrates are added to the synthesized proteins?

Rough Endoplasmic Reticulum

What are telomeres?

They are multiple repetitive nucleotide sequences at the ends of chromosomes.

Histone is a _________________.

DNA binding protein

Which restriction enzyme did Herb Boyer discover in 1970?

EcoRI

What enzyme separates strands of DNA during DNA replication to make DNA single-stranded so it can be copied?

DNA helicase

During DNA replication, the enzyme ______________is involved in unwinding the DNA double helix at the replication fork, while ______________ catalyzes the addition of nucleotides to the 3’ end to generate new polynucleotide strands.

Helicase: DNA polymerase III

T/F: DNA polymerase III removes the RNA primers and replaces them with DNA.

False

T/F: Adenine forms 3 hydrogen bonds with thymine and guanine forms 2 hydrogen bonds with cytosine

False

T/F: The leading strand is replicated continuously, while the lagging strand is replicated discontinuously

True

T/F: Chromatin is a complex of DNA and fatty acids

False

T/F: A DNA strand has polarity with a phosphate group in the 5’ end, and a OH group in the 3’ end

True

T/F: DNA replication is semi-conservative; the resulting daughter DNAs will each consist of a parent strand and a newly synthesized strand.

True

T/F: New DNA is synthesized in a 5' to 3' direction

True

Nonprotein coding pieces of pre-mRNA that are removed during RNA splicing are called ________.

Introns

Which of the following is a polyadenylation signal sequence of mRNA.

AAUAAA

What is a polyadenylation signal sequence?

It's a poly(A) tail in mRNA that adenine bases.

What is the difference between transcription and translation?

Transcription is the process of synthesizing RNA from the DNA template strand which occurs inside the nucleus, while translation is the process by which RNA is translated into amino acids which then form into protein. This occurs outside of the nucleus.

Addition of a poly(A) tail to an mRNA molecule ________.

Allows mRNA molecules to be more stable in the cytoplasm

In eukaryotes, promoters include a _________ box, which is crucial for the formation of the transcription initiation complex.

TATA

Which type of RNA molecule contains the genetic code of a gene that is read by ribosomes during translation?

mRNA

Which enzyme is involved in "transcription".

RNA Polymerase

Which of the following takes the genetic code from the nucleus to the cytoplasm?

mRNA

T/F: In eukaryotes, the primary mRNA is not functional, and modified by RNA processing to form mature mRNA.

True

Which of the following RNA modifications could best be described as "removal of intron sequences"?

Splicing

Which is the starting codon in protein translation?

AUG

What are restriction sites?

Restriction sites are the specific palindromic sequences from which restriction enzymes cut DNA.

Alpha helices and β-pleated sheets represent the ________ structure of a protein.

Secondary

is a gene found in cloning vectors that is used for easy screening of bacteria that carry the recombinant plasmids as opposed to non-recombinant plasmids, on plates containing X-Gal.

B-galactosidase gene

T/F: Accurate translation can only occur when ribosomes examine codons in-frame.

True

A point mutation that changes a codon specifying an amino acid into a stop codon is called a________.

Nonsense mutation

A mutation where a base changes, but the same amino acid is encoded is called a__________.

Silent mutation

A mutation in which a different amino acid is encoded that may or may not affect protein function is called_________.

Missense Mutation

A mutation involving insertions or deletions are called __________.

translational frameshift mutations

T/F: A restriction enzyme generally recognizes and cuts many different combinations of nucleotide sequences

False

A bacterial cell that is able to take up naked or recombinant DNA is said to be___________________.

Competent

Transformation in a cloning experiment is ________.

Inserting recombinant plasmid into bacterial cells.

In the given DNA nucleotide sequence, 5’ AATGGCA 3’ . What is the +2 reading frame?

5’ ATGGCA 3

What is a palindrome?

A sequence wherein reading 5' to 3' forward on one strand matches the sequence reading backward 5' to 3' on the complementary strand.

Ex:

5' GAATTC 3'

3' CTTAAG 5'

Quaternary structure of a protein refers to ________.

The association of multiple polypeptide subunits to form a functional protein

Plasmids can best be described as:

Small, circular DNA molecules that can exist independently of chromosomes commonly found in in bacteria.

________________ expression system does not carry the animal virus.

Plant

______________________ is the generation of multiple identical copies of a gene or defined DNA segment.

DNA cloning

Adding a phosphate or sugar group to a newly made protein is an example of ________.

A posttranslational modification

Northern hybridization is a technique for the ____________?

Detection of specific RNA fragments separated by gel electrophoresis

Which one of the following Next Generation Sequencing techniques provides less error?

Illumina's sequencing by synthesis

In the polymerase chain reaction (PCR) technique, there is a heating phase and an alternate cooling phase. An original sample of DNA would have to pass through how many total rounds of heating and cooling before a sample is increased eight times in quantity?

Three

The human genome is thought to contain about how many genes?

45,000

RNA is copied into complementary DNA by ­­­­­­­­­­­­­­­­­­­­­­­­­­______________.

Reverse transcriptase

How many proteins are approximately encoded from human genes?

More than 120,000

Which of the following technique can be used to transfer DNA onto nitrocellulose membrane?

Southern blotting

MALDI-TOF-MS is ________.

Matrix-assisted laser desorption/ionization - time of flight - mass spectrometer

In Sanger’s original dideoxy chain termination DNA sequencing technique, DNA polymerase stops adding a new dNTP because the last molecule added is lacking:

A 3' OH group on the ribose sugar

Imagine you wanted to use a human genomic DNA library to clone the human gene for insulin. You will be using the rat insulin gene sequence as your DNA probe. In what order would you perform the following steps to accomplish this goal?

1. Use autoradiography to identify colonies containing DNA that hybridized to the probe

2. Grow transformed cells on media with antibiotics and X-gal for blue-white screening

3. Ligate genomic DNA and vector DNA

4. Cut genomic DNA and vector DNA with restriction enzymes

5. Hybridize library DNA with labeled probe for the rat insulin gene

6. Transform bacteria with recombinant plasmid

4, 3, 6, 2, 5, 1

You forget to include sodium dodecyl sulfate (SDS) in your samples for SDS-PAGE. What happens when you run the gel?

The proteins would not migrate towards the positive electrode.

What is SDS?

sodium dodecyl sulfate is a detergent used in Polyacrylamide Gel Electrophoresis (PAGE).

Gives proteins a negative charge so that they will migrate towards the positive electrode.

Describe FISH

Stands for FluorescentIn Situ Hybridization. Uses f luorescent dyes attached to probes to identify the locationof specific mRNAs in place in intacttissues or organisms.

What is Southwestern blotting?

Involves identifying and characterizing DNA-binding proteins (proteins that bind to DNA).

Which is the starting codon in nucleotide sequence?

ATG

In a recombinant DNA experiment, which enzyme covalently repairs the phosphodiester bond between the 3’ OH end of one DNA fragment and the 5’ PO₄ ^3- end of another DNA fragment?

DNA ligase

Which of the following restriction enzyme is isolated from the bacterium Bacillus amyloliquefaciens?

Bam HI

Compared with traditional biotechnology, modern biotechnology is characterized by______________________________ technique.

Genetic Engineering

A bacteriophage is ________.

A virus that specifically infects bacteria

gene encodes the enzyme B-galactosidase that catalyzes the hydrolysis of X-Gal.

LacZ

A ________ is a single-stranded DNA molecule attached to a radioactive or fluorescent compound that is complementary to a specific sequence of DNA. Such pieces of DNA are used to identify and study cloned genes in hybridization experiments.

Probe

The entire genetic code consists of ____ amino acids.

20

Which of the enzymes recognition sites would produce sticky ends compatible with (5'-G|AATTC-3')?

EcoRI (3'-CTTAA|G-5')

During library screening, PCR, Southern blotting, and other techniques, binding two pieces of DNA to each other by hydrogen bonding is called ________.

Hybridization

Which promoter is commonly used for protein expression in plants?

CaMV35S

Dideoxyribonucleotides (ddNTPs) used for DNA sequencing lack oxygen atoms at ________.

The 3' carbon of the pentose sugar

Which one of these protein analysis techniques do not involve immunoassays?

MALDI-TOF-MS

In DNA, which of the base pairs have three hydrogen bond?

C - G

In the given DNA nucleotide sequence, 5’ AATGGCA 3’ . What is the +3 reading frame?

5’ TGGCA 3’

During rapid growth, the cell membrane of E. coli has hundreds of pores, called __________.

adhesion zones

Define TATA box

a DNA sequence that indicates where a genetic sequence can be read and decoded. It is a type of promoter sequence, which specifies to other molecules where transcription begins.