Human Genome Project

-to sequence all DNA in the Human Genome

-to identify location and function of every gene

-2003 99% sequenced

-hundreds of disease associated genes Id'd

Recombinant DNA Technology

-lab techniques for combining genes from different sources & species, into single DNA molecule

-use of recombinant DNA to produce useful products

Plasmids

a genetic structure in a cell that can replicate independently of the chromosomes, typically a small circular DNA strand in the cytoplasm of a bacterium or protozoan. Plasmids are much used in the laboratory manipulation of genes.

Cloning and 2 types

-Reproductive Cloning- results in the birth or a new organism

-Therapeutic Cloning- can make cells and tissues for medical use (cloning of the dna or gene)

6 Steps in Cloning

1. Get an egg cell from an animal

2. Remove the nucleus

3. Add a body cell from an adult donor to the empty cell

4. Grow in culture to produce a blastocyst

5. Implant embryo in surrogate mother(or implant in culture)

6. CLone of donor is born

Restriction Enzyme

-the cutting tool.  Recognizes short nucleotide sequences in DNA and cuts a specific points

-cuts into sticky and blunt ends

Transformation

-the taking up of DNA from the fluid surrounding the cell

ex. Griffith - a harmless strain of bacteria took up pieces of DNA left from the dead cells of a disease-causing strain

Transduction

-the transfer of bacterial genes by a phage

-the phage has a fragment of DNA from its host cell (a stowaway)

Conjugation

-a "male" bacterial cell attatches to a "female" cell by sex pilli, a bridge forms, and DNA passes from the male to female

Gel Electrophoresis

-sorts macromolecules (proteins or nucleic acids) by their charge and length

-Each DNA mixture is placed into a well in a gel

-A negative electrode is attached to gel at DNA end, and positive electrode to the other end

-DNA fragments have a negative charge due to (PO4-) groups, & move towards posative pole. Shorter fragments move faster than longer ones

Polymerase Chain Reaction (PCR)

-cloning/ampliphying DNA

-DNA is heated, mixed with DNA polymerase, nucletide monomers & short pieces of synthetic single-stranded DNA

-DNA is replicated into 2 daughter DNA's

-fast

Sticky and Blunt Ends

S: double stranded with single stranded part at end, key to joining with other DNA

-join with a DNA from another source, cut with same restriction enzyme with 1st DNA

B: used for forensic sciene, gel electrophorysis, in the lab we did the rectangles of DNA

DNA Technology and Ethical Concerns

DT- methods to study and manipulate genes

EC- create pathogens, organisms cannot survive out of lab, foods? ex. superweeds, must be identified

Steps in Gene Therapy and Why

1. normal human gene is isolated an cloned

2.human gene is inserted un the virus/vector

3. the virus is injected into the patient

-could correct a genetic disorder or treat a medical problem

Steps for Making a Protein

1. Cell containing gene of interest

2. Recombinant DNA (host DNA plus gene of interest

3. Cell multiplies and produces proteins

4. Genes may be inserted into another organsim or proteins may be harvested

Splicing DNA into a Plasmid

1. Restriction enzyme cuts DNA into fragments

2. Addition of DNA fragment from another source

3. Fragments stick together by base pairing, DNA ligase connects the DNA pieces

DNA Fingerprinting

-a procedure that analyzes a persons unique collection of DNA restriction fragments

-used in forensic science

-determines whether 2 samples of DNA are from the same individual