DNA Tech-Genomics-Proteomics

Question 1

Biotechnology

Answer 1

any technique applied to biological systems to manipulate processes for a specific purpose

Question 2

DNA technologies

Answer 2

isolate, purify, analyze and manipulate DNA sequences

Question 3

Genetic engineering

Answer 3

uses DNA technologies to alter genes for practical purposes

Question 4

Cloning DNA

Answer 4

break open cells, let chromosomes come out, get the plasmid out of bacteria and cut open the plasmid using restriction enzymes (the same restriction enzyme to cut DNA and to open plasmid), use DNA ligase to attach the pieces of DNA with a plasmid

Question 5

Bacterial plasmids that are engineered to contain:

Answer 5

1. gene of interest

2. sorting genes

Question 6

How can we isolate the bacteria that has our gene of interest?

Answer 6

Make agar plate and add ampocilin, if the bacteria doesn’t have the AMP gene it will die. Add X-gal substrate for the enzyme beta-galaticylase, if beta breaks down X-gal it turns blue (meaning it has the gene), even if it has the gene there can not be a gap in between

Question 7

How to further isolate the gene of interest?

Answer 7

use a nucleic acid probe to identify gene of interest in set of clones. Break open the cells and denature them (make single stranded), add a single stranded DNA probe that only matches the gene of interest and add some form of dye. Observe which pot the gene is in

Question 8

Genomic library

Answer 8

collection of clones containing every sequence in a genome

Question 9

Complementary DNA (cDNA) library

Answer 9

DNA sequences made from expressed RNA

Question 10

Make cDNA

Answer 10

go back into cell, build a complementary DNA strand using reverse transcriptase, add a complementary strand onto mRNA, every single mRNA has a polyAtail so that we have the primer and reverse transcriptase. Get rid of the RNA strand (using enzymes) and add DNA polymerase to do DNA replication

Question 11

Polymerase chain reaction

Answer 11

denaturation, annealing, replication, taq polymerase. Denature DNA by increasing temp. add a DNA polymerase and nucleotides to start replicating DNA. Need to add taq polymerase to withstand. Repeat at least 30 times

Question 12

Restriction fragments length polymorophisms

Answer 12

different fragments from the same part of genome

Question 13

Southern blot analysis

Answer 13

uses electrophoresis, blot transfer, and labelled probes to identify RFLPs

Question 14

DNA fingerprinting

Answer 14

observing short tandem repeats

-parts of nucleotide sequence that is repeated over and over

Question 15

Transgenic organisms

Answer 15

modified to contain genes (trangene) from external source

Question 16

Animal genetic engineering

Answer 16

use germ-line insert transgene, allow to grow and divide, inject into an embryo, place in a surrogate mouse. Some will have transgene, some wont

Question 17

Stem Cell

Answer 17

unspecialized cell that can:

• reproduce indefinitely
• differentiate into various specialized cells

Question 18

Embryonic stem cells

Answer 18

isolated from early embryos at the blastocyst stage

-can differentiate into any cell types (pluripotent)

Question 19

Adult stem cells

Answer 19

replace nonreproducing specialized cells

Question 20

Gene therapy

Answer 20

alteration of individual’s genes. May treat disorders with one defective gene

Question 21

Dolly

Answer 21

take a cell from mammory gland, get an egg from another ewe and remove the nucleus and fuse the new nucleus into the egg and implant it into a third sheep as a surrogate

Question 22

Genomics

Answer 22

is the characterization of whole genomes (structure, function, evolution)

Question 23

Sequencing

Answer 23

determine the nucleotide sequence of a genome

Question 24

Annotation

Answer 24

analyses specific regions of nucleotide regions

Question 25

Functional genomics

Answer 25

studies functions of genes and other parts of genome

Question 26

Comparative genomics

Answer 26

studies how genomes have evolved by comparing genomes of various organisms

Question 27

Dideoxy (Sanger) method

Answer 27

• dideoxyribonucleotides have -H bound to 3’ C instead of -OH
• DNA polymerases place dideoribonucleotides in DNA, stops replication
• polyacrylamide gel separates strands varying by one nucleotide

Question 28

Whole-genome shotgun method

Answer 28

DNA purification, DNA fragmentation, amplificaiton of fragments, sequencing each fragment, assembly of fragments into genome sequences

Question 29

Human genome project

Answer 29

took 13 years (2003)

• revolutionizing biology and evolutionary understanding
• 3.2 billion base pairs
• between 20000 and 25000 protein coding genes
• protein coding 2% of genome

Question 30

Transcriptome

Answer 30

complete set of transcripts

Question 31

Transciptomics

Answer 31

study of transcriptome

Question 32

Proteome

Answer 32

• complete set of proteins expressed by genome

- larger than genome in eukaryotes

Question 33

Proteomics

Answer 33

study of proteome to determine:

-structure, location, interactions, functions

Question 34

Interactome

Answer 34

sum of all interactions of all gene products in a cell