Biotechnology

30
Biotechnology repairstemcell.files.wordpress.com Dolly

description

Biotechnology. Dolly. repairstemcell.files.wordpress.com. Potential Uses for Cloned Genes. to produce a protein product plasminogen activator to endow an organism with a metabolic capability engineered bacteria that degrade oil spills create more copies of the gene for further study. - PowerPoint PPT Presentation

Transcript of Biotechnology

Page 1: Biotechnology

Biotechnology

repairstemcell.files.wordpress.com

Dolly

Page 2: Biotechnology

Potential Uses for Cloned Genes to produce a protein product

plasminogen activator

to endow an organism with a metabolic capability engineered bacteria that degrade oil spills

create more copies of the gene for further study

Page 3: Biotechnology

Genetic Engineering Stages

1. Isolate gene cleavage of plasmid 2. Recombinant-DNA 3. Clone * Screen for quality 4. Harvest

Gene Protein

Page 4: Biotechnology

1. Isolate Gene

Isolate the desired gene from the genome

the desired gene will be cut with restriction endonucleases (enzymes)

desired gene to beisolated

Page 5: Biotechnology

Cleavage

cut DNA of host with restriction endonucleases (enzymes)

several hundred RE’s exist discovered 1960s

produce sticky ends or blunt ends

Page 6: Biotechnology

Sticky Ends

Escherichia coli

Eco RI

sticky end

Page 7: Biotechnology

Blunt ends

Haemophilus aegyptius Hae III

G G C C

C C G G

G G C C

C C G G

Page 8: Biotechnology

2. Recombinant DNA

splice desired gene into host DNA, DNA ligase seals the strands

Vector, generally plasmid of bacteria (prok) or yeast (euk) viral DNA (not for harvesting protein)

plasmid with recombinant DNA

Page 9: Biotechnology

3. Clone

produce a cell line in which all members have identical copies of a particular gene

Screen Choose cells that carry desired gene & eliminate those cells that do not carry desired gene

Page 10: Biotechnology

4. Harvest (or Isolate)

harvest protein harvest gene

genetic harvesting protein harvesting

pest resistant gene oil eating bacteria dissolving clot protein human growth hormone

copies of the gene

protein molecules

Page 11: Biotechnology

Cloning

Directly from an organism complementary DNA

made from mRNA template through

reverse transcription (cDNA) Reverse transcriptase can be used

to make smaller cDNA libraries These contain only the genes that are

transcribed by a particular type of cell recognized by the addition of a RE

recognition sequence to it

reversetranscriptase plus mRNAs

mRNAs

mRNA is degradedby an enzyme

DNA polymerasesynthesizes the 2ndstrand

cDNA

Page 12: Biotechnology

Genomic Libraries

“Book,” a clone containing a foreign DNA

Plasmid library (bacterial, yeast) Phage library (virus) Bacterial Artificial Chromosome library

Page 13: Biotechnology

Plasmid Library

Copies of DNA fragments can be stored in a cloned bacterial plasmid

Each one of these is considered a "book"

recombinant plasmid

bacterial clones

foreign genome

Page 14: Biotechnology

Phage Library

DNA fragments can be stored in a cloned phage

each phage type isconsidered a "book"

phage clones

recombinant phage "book"

foreign genome

Page 15: Biotechnology

Bacterial Artificial Chromosome (BAC) LibraryCopies of multiple

DNA fragments can

be stored in a bacterial

artificial chromosome plasmid with many genes

BACclone

each clone occupies one well

Page 16: Biotechnology

Gene products Product Made in Use human insulin E. coli diabeteshuman growth hormone E. coli growth defectsepidermal growth factor E. coli burns, ulcersinterleukin-2 E. coli possibly cancerbovine growth hormone E. coli improving weight gaincellulase E. coli breakdown of celluloseTaxol E. coli ovarian cancerhepatitis B vaccine S. cerevisiae prevents hepatitiserythropoietin mammalian cells anemiafactor VIII mammalian cells hemophiliatissue plasminogen activator mammalian cells heart

attacks

Page 17: Biotechnology

Other Examples

"golden rice" genetically modified rich in beta-carotene prevents blindness

news.bbc.co.uk

Page 18: Biotechnology

papaya's ring spot disease gene was introduced to

control the plague

http://www2.dpi.qld.gov.au

Page 19: Biotechnology

Human Genome Project

Collaborative effort to map and sequence entire human genome

Began 1990 4 goals

genetic (linkage) mapping physical mapping sequencing analyzing the genomes of other species

Page 20: Biotechnology

Genetic Mapping of the Human Genome to locate genetic markers spaced evenly

throughout the chromosomes

to make it easier to find other loci

Page 21: Biotechnology

Physical Mapping of the Human Genome cutting chromosomes into identifiable

fragments then determining their order on the chromosome

Page 22: Biotechnology

Sequencing the Human Genome determining the exact nucleotide pairs haploid set of human chromosomes contains

approximately 3 billion nucleotide pairs Genbank

Database where DNA sequences have been deposited publicly available via the Internet

final draft, 2004 (over 99% of genome was determined)

remain a few 100 gaps of unknown sequences that require special methods to figure out

Page 23: Biotechnology

Analyzing Gene Expression Analyze genomes of other important species

for genetic engineering

Page 24: Biotechnology

Stem Cells

unspecialized blastula cells pluripotent

adult stem cells gives rise to specific types of cells bone marrow blood cells

Page 25: Biotechnology

Applications Medical

Diagnosis Human Gene therapy Pharmaceutical products

animal and plant application gold rice salinity resistant gene

Environmental biofuel oil cleaning bacteria

Forensic evidence The Innocence Project conviction of guilty

Page 26: Biotechnology

Genomes of other species and H. sapiens Bacteria

H. influenzae 1,700 1995 E. coli 4,400 1997

Fungi S. cerevisiae 6,200 1996

Plants Oryza sativa (rice) 60,000 2002

Animals D. melanogaster 13,700 2000 Mus musculus 22,000 2001 Rattus norvegius 25,000 2004 H. sapiens 21,000 2003

Page 27: Biotechnology

Ethical Issues

Should we engineer new genotypes for individuals with anomalies? diabetes, CF, immune deficiencies, MD, stunted

growth, sickle-cell disease myopia, altering personalities, increase length of

life

Page 28: Biotechnology

Should we engineer human germ cells? If they are carrying abnormal genes

eugenics - deliberate effort to control the genetic makeup of human populations color of eyes color of skin color of hair

Page 29: Biotechnology

We have technology to test for diseases for which there is no cure and sometimes no treatment. (Ex. Huntington’s disease, breast cancer)

Would you want to be tested?

Page 30: Biotechnology

Who should have right to examine someone’s genetic info?

How should that info be used? Should a person’s genome be a factor in

determining eligibility for a job or insurance

The End