Biotechnology

Question 1

DNA Technology

Answer 1

techniques to isolate, purify, analyze, and manipulate DNA sequences

Question 2

Genetically modified organism (GMO)

Answer 2

Genetically altered organism whose genome has been engineered to introduce or change a genetically controlled trait

Question 3

Biotechnology

Answer 3

any technique used to make or modify products or processes
of a biological system or living organism

Question 4

Gene cloning

Answer 4

the method of producing many identical copies of a gene of interest in a host cell

Question 4

Restriction Endonuclease

Answer 4

Recognizes specific DNA sequences called restriction sites, cleaves them, and creates what’s called restriction fragments - ends are sticky

Question 5

The plasmid interacting with the gene of interest

Answer 5

The gene of interest is placed within the Plasmid, and ligase is used in order to seal the nick in the sugar backbone - after this process, the plasmid is now a Recombinant Plasmid

Question 6

Identifying the Recombinant Plasmid with Gene of Interest

Answer 6

Recombinant Plasmid is then placed in a solution with E coli bacteria - some of the bacteria pick up the plasmid and some don’t.

The recombinant plasmid and normal plasmids are placed on a plate in which a antibiotic is introduced, because the recombinant plasmid contained a gene that is resistant to it, the normal plasmid die off (these ones are blue)

Question 7

Polymerase Chain Reaction (PCR)

Answer 7

making a lot of copies of the same DNA sequence

Question 8

Process of PCR: 3 steps

Answer 8

DNA strands are DENATURED through heating (95c)

ANNEALING: primers are placed on the DNA strands in a complementary sequence,

Extension: sample is then heated to 72C, DNA polymerase 1 is then used to extend the primers and make 2 identical copies of DNA

After each cycle, the DNAs are doubled consistently

Question 9

Gel Electrophoresis

Answer 9

Used to separate DNA, RNA, and other protein molecules

separates by size, charge, or other properties, DNA strands are smaller go to the bottom and the ones that are larger stay at the top.

This is because larger strands are more negative, and smaller ones aren’t as negative, so they’re organized by size as well as charge.

Question 10

Agarose gel electrophoresis

Answer 10

The gel the molecules are placed in, used to compare the size of the DNA strands, commonly used for DNA fragment analysis

Question 11

Central Dogma of Biology:

Answer 11

DNA → RNA → Protein

Question 12

Reverse transcriptase is..

Answer 12

the only enzyme that disobeys the central dogma

Question 13

Process of making cDNA

Answer 13

mRNA is isolated and has the poly-A tail attached to it

Question 14

What primer is placed on the mRNA?

Answer 14

oligo (DT) which is just a strand of T, complementary to the poly-A Tail

Question 15

After the oligo (DT) is placed

Answer 15

DNA Transcriptase synthesizes the complementary strand

as a result we now have a mRNA strand as well as the cDNA strand

Question 16

How is the mRNA removed from the single strand of cDNA

Answer 16

DNA polymerase type 1 kicks out some strands of RNA leaving nicks in the between

Then it Synthesizes the commentary strands, while also removing the remainder of the rest of the mRNA strands - ligase is then used to finish the job

Question 17

What is an expression vector and what is it used for?

Answer 17

An expression vector is a cloning vector that is very similar to the plasmid in that it allows for the expression of a gene of interest.

Question 18

Transgenic

Answer 18

Organisms that have had their genes altered

Question 19

Gene Targetting

Answer 19

Knocking out, Replacing, or adding genes into a genome

Question 20

Stem cells

Answer 20

cell that are undifferentiated, that do not have a specific job, they have the chance to of becoming other cells in the body and also replace worn out cells when they die

Question 21

Gene Therapy

Answer 21

The introduction of healthy gene into a cell line to correct a genetic disorder

Question 21

Embryonic Stem Cells: pluripotent

Answer 21

These cells are only found in early stage embryos and are pluripotent, meaning they have the ability to differentiate into any cell line - any tissue

Question 22

Adult stem cells: multipotent

Answer 22

are tissue specific and function to replace specialized cells of tissues that are multipotent. Meaning they are limited in the amounts of cells they can give rise to

Question 23

What are knockout mice and how are they created?

Answer 23

Knock out mice are organisms that have had their genes altered and have turned them nonfunctional.

Embryonic stem cells are mice are extracted, introduced to transgenes and then injected back into early-stage embryonic mice. This is done in order to figure out the function of a specific gene and getting rid of the gene is the best way to do so.

The reasoning for targeting the embryonic cells is because those are the ones that produce the rest of the cells in the body so when they multiple there is no expression of that gene. Mice with the genetic change are bred and the homozygous recessive mice are the ones that are the knockout mice.

Question 24

CRISPR - Cas 9

Answer 24

A virus injected into the cell, the CAS 9 protein has a guide RNA within it that codes for the genetic change.

CAS 9 opens the DNA strand and looks for the PAM sequence that it will then cut. It specifically cuts at the PAM sequence because that is where the guide RNA will be placed, DNA poly is then used to synthesize the rest of the DNA.

Question 25

Germline Therapy

Answer 25

When the gene is introduced in germline of the human - the sperm of the human so that their offspring do not have the genetic mutation - however this is not allowed in humans

Question 26

Somatic Gene Therapy

Answer 26

Adult body cells are cultured and transformed with an expression vector containing target transgene and re introduced to the body - allowed in humans

Question 27

Sickle Cell Mutation and Restriction Fragment Length Polymorphisms

Answer 27

Caused by a single base pair mutation - one nucleotide change, restriction fragment length polymorphisms are shown through the electrophoresis in which the length of restriction sites are different

Question 28

DNA Fingerprinting

Answer 28

PCR is used to analyze specific DNA sequence variations at specific loci in the genome which is then analyzed by the gel electrophoresis

Question 29

Short Tandem Repeats

Answer 29

Short Sequence of DNA repeated in a sequence, the number of repeats at each loci varies among individuals in the population