Bio Chapter 20

Front 1

Genome

Back 1

organization of all of the genes in an organism

Front 2

Biotechnology Industry

Back 2

diagnosis of diseases, gene therapy, vaccines/pharmaceutical products, forensic uses, environmental/agricultural uses

Front 3

Human Genome Project

Back 3

1) Map all genes in human genome
2) Sequence all genes in human genome
3) Map & sequence model genomes (yeast, mice)
4) ELSI - ethical, legal, and social implications

Front 4

Screening for: Parents, Sperm, Embryos etc., later in life

Back 4

Parents: Carrier status
Sperm: Sex of child
Embryos etc.: genetic disorders
Later in life: predispositions to disease, life planning

Front 5

Gel electrophoresis

Back 5

Technique which separates nucleic acid or proteins on the basis of size, electrical charge, and other physical properties

Front 6

Dr. Alec Jeffreys

Back 6

Developed DNA profiling procedure

Front 7

Restriction Enzymes

Back 7

Bacterial enzymes that recognize short DNA nucleotide sequences
DNA pieces = restriction fragments

Front 8

Gel electrophoresis Procedure

Back 8

1) Isolate pure DNA
2) Cut DNA w/ restriction enzymes
3) Load DNA fragments into wells
4) Use gel electrophoresis to separate DNA fragments by size

Front 9

4 materials of gel electrophoresis & what they do

Back 9

Electricity: propel DNA through agarose gel
Agarose gel: acts like a strainer to separate by size
Well: force DNA to move through agarose
Control DNA: DNA fragments of known size

Front 10

RFLP Analysis

Back 10

Procedure to highlight minute differences in human individual DNA by tagging (99% alike)

Front 11

RFLP Analysis Procedure

Back 11

1) Restriction enzyme digestion of DNA
2) Gel electrophoresis
3) Southern Blot
4) Autoradiography: radioactive probes expose film & create DNA bond image

Front 12

Southern Blot

Back 12

1) Denature DNA (single stranded)
2) Transfer denatured DNA to nylon membrane
3) Add radioactive probes (small single stranded DNA complementary to DNA of interest)
4) Probes bind

Front 13

Applications of RFLP Analysis

Back 13

-Diagnose disease/carriers/prenatal samples
-Prepare fingerprints
-Compare genomes (different species/evolution)
-Paternity/Maternity
-Individual identity
-Lineage

Front 14

PCR Requirements

Back 14

DNA w/ sequence of interest, special heat resistant DNA polymerase, nucleotides, artificial primers (complementary to ends of targeted sequence)

Front 15

PCR Procedure

Back 15

1) Add heat to denature DNA
2) Add artificial primers
3) Cool to allow primers to anneal (stick)
4) Add taq polymerase (heat resistant) to extend chain
5) Repeat cycle, creates chain reaction to expedite copying of DNA of interest

Front 16

PCR Advantages/Disadvantages/Applications

Back 16

Advantage: faster than gene cloning using bacteria
Disadvantage: errors accumulate over many copies
Applications: amplify DNA of very small samples

Front 17

Genetic Engineering (Gene Cloning)

Back 17

Direct manipulation of DNA for practical purposes
Goal: to get host to treat recombinant DNA as natural part of its genome

Front 18

Genetic Engineering Tools

Back 18

Restriction Enzymes (scissors): create sticky ends
Ligase (glue): creates sugar phosphate bonds
Carrier of DNA (vector): virus, plasmid, engineered retrovirus
Host organism

Front 19

Genetic Engineering Procedure

Back 19

1) Isolate plasmid & gene of interest
2) Cut w/ same restriction enzyme
3) Mix plasmid w/ human gene
4) Add ligase (seal SP bonds)
5) Transformation
6) Binary Fission

Front 20

Recombinant DNA

Back 20

DNA that contains genes from two different sources

Front 21

Ensuring Expression of Gene of Interest in Recombinant DNA

Back 21

Produce cDNA (complementary)
1) isolate mRNA that has already been processed
2) use reverse transcriptase from retrovirus to make cDNA

Front 22

Applications of Genetic ENgineering

Back 22

1) Manufacture of human proteins
2) Creating transgenic organisms (GMOs)
3) Gene therapy

Front 23

DNA (Genomic) Libraries

Back 23

1) Genomic library: contains cloned genes from the entire genome of an organism
2) cDNA library: all mRNAs collected from a cell, and reverse transcribed into cDNA

Front 24

DNA Microarray Assay (DNA chip)

Back 24

Single stranded DNA (usually genes) fixed to wells in a glass slide
-Genetic Predisposition Profile

Front 25

Automated DNA Sequencing

Back 25

Decoding linear order of the bases

Front 26

Cloning

Back 26

-Nuclear transportation
-High incidence of disease mutation
Ultimate goal: supply cells for repair of damaged/diseased organs (therapeutic cloning)

Front 27

Stem Cells

Back 27

1) Embryonic- blastocyst stage, pluripotent (differentiate into many different types of cells
2) Adult- can be reprogrammed

Front 28

Genome Evolution

Back 28

-Alterations of chromosome structure
-Duplication & divergence of genes
-Homeotic (developmental) genes contain a "homebox" region whose sequence is highly conserved amongst diverse species