| front |
back |
revisions |
lasted changed by |
history |
| Recombinant DNA technology |
making recombinant DNA involves three processes:
1. manipulation of DNA invitro (that is out side cells)
2. VECTOR. the recombination of another organisms DNA with bacterial DNA in a phage or a plasmid.
3. the cloning, or production of many genetically identical progent, of phages or plasmids that carry foreign DNA.
2 DNA's put together.
You have to have one gene of interest, one plasmid or phage, and one bacteria. |
0 |
WilkieLW Thu, 11 Mar 2010 20:10:40 GMT |
 |
| Bacteria are commonly used as hosts |
b/c if easily isolation * DNA bacterial cultures grow quickly >rapidly cloning the instered forgeign gene. |
0 |
WilkieLW Thu, 11 Mar 2010 20:10:40 GMT |
|
| Vectors |
(bacterial plasmid or virus) are used for moving DNA from test tubes-> into cells. |
0 |
WilkieLW Thu, 11 Mar 2010 20:10:40 GMT |
|
| Restriction enzymes |
cut DNA at specific nucleotide sequences. Instead of cutting the molecules straight accross, they leave "sticky" ends. Any DNA cleaved by such and enzyme can be joined readily to another DNA molecule cleaved by the same enzyme. (**DNA ligaseis used for permanent union (2 dif DNA's)
endonuclease |
1 |
WilkieLW Thu, 11 Mar 2010 20:12:04 GMT |
|
| Recombinant DNA technology |
refers to the set of technologies for recombinating genes from different source in vitro and transferring this recombinant DNA into a cell (invitro) where it may be expressed. (translate into MRNA to protien)
>includes methods for:
1. obtaining DNA segments short enough to be analyzed and manipulated.
2. obtaining large quantities of identical DNA segments.
3. Determining the exact order of the nucleotides in a DN segement.
4. Locating and identifying specific DNA segments of interest. |
0 |
WilkieLW Thu, 11 Mar 2010 20:00:50 GMT |
|
| DNA technology |
is a powerful set of technologies that help to manipulate and analyse genetic material.
TOOL KIT--->
1. restriction enzymes and other enzymes
2. DNA vectors
3. Host organisms. |
0 |
WilkieLW Thu, 11 Mar 2010 20:00:50 GMT |
|
Genetic Engineering
Antibiotics |
inhibit or kill microbial growth, (increase yeilds by gene amplification.) |
0 |
WilkieLW Thu, 11 Mar 2010 20:00:50 GMT |
|
Genetic Engineering
Human hemoglobin |
blood substitute in emergencies (produced in gene-altered pigs) |
0 |
WilkieLW Thu, 11 Mar 2010 19:53:02 GMT |
|
Genetic Engineering
AIDS subunit vaccine (in clinical trials) |
incomplete virus vaccine |
0 |
WilkieLW Thu, 11 Mar 2010 19:53:02 GMT |
|
Genetic Engineering
Hepatitis A and B vaccines |
Prevent Hepatitis |
0 |
WilkieLW Thu, 11 Mar 2010 19:53:02 GMT |
|
Genetic Engineering
Epidermal growth factor |
heal wounds |
0 |
WilkieLW Thu, 11 Mar 2010 19:53:02 GMT |
|
Genetic Engineering
Bone growth factor |
heal bone fractures, treat osteoporosis, stimulate bone growth. |
0 |
WilkieLW Thu, 11 Mar 2010 19:51:23 GMT |
|
Genetic Engineering
Erythropoietin |
treat anemia, stimulate formation of new red blood cells. |
0 |
WilkieLW Thu, 11 Mar 2010 19:51:23 GMT |
|
Genetic Engineering
Blood clotting factor VIII |
treat hemophilia |
0 |
WilkieLW Thu, 11 Mar 2010 19:51:23 GMT |
|
Genetic Engineering
Human growth hormone |
prevent pituitary dwarfism |
0 |
WilkieLW Thu, 11 Mar 2010 19:51:23 GMT |
|
Genetic engineering
Human Insulin |
Treats diabetes |
0 |
WilkieLW Thu, 11 Mar 2010 19:49:33 GMT |
|
| Genetic engineering |
refers to the purposeful manipulation of genetic material to alter the characteristics of an organism in a desired way. (fruits and veg, medication) |
0 |
WilkieLW Thu, 11 Mar 2010 19:49:33 GMT |
|
| Transposon |
is a transposable element that contains the genes for transposition. |
0 |
WilkieLW Thu, 11 Mar 2010 19:49:33 GMT |
|
| Transposable Element |
mobile genetic sequence. The simplest type of transposbale element, an insertion sequence. |
0 |
WilkieLW Thu, 11 Mar 2010 19:49:32 GMT |
|
| Transposition |
the ability of a genetic sequence to move from one location to another. |
0 |
WilkieLW Thu, 11 Mar 2010 19:44:47 GMT |
|
| Table 8.2 Page 222. |
. |
0 |
WilkieLW Thu, 11 Mar 2010 19:44:47 GMT |
|
| Catabolic Enzymes |
some plasmids contain genes for catabolic enzymes. Generally, plasmids carry genes that code for functions not essential for cell growth, the chromosome carries the genes that code for essential functions. |
0 |
WilkieLW Thu, 11 Mar 2010 19:44:47 GMT |
|
| Tumor-inducing (Ti) plasmids |
can cause tumor formation in plants (affect plants) |
0 |
WilkieLW Thu, 11 Mar 2010 19:44:47 GMT |
|
| Virulence plasmids |
such as those in salmonella, or the neurotoxin genes carried on plasmids in Clostridium tetani, cause disease signs and symptoms |
0 |
WilkieLW Thu, 11 Mar 2010 19:39:49 GMT |
|
| Other plasmids |
direct the synthesis of bacteriocidal (bacteria-killing) proteins called bacteriocins. |
0 |
WilkieLW Thu, 11 Mar 2010 19:39:49 GMT |
|
| Resistance (R) plasmids |
carry genes that provide resistance to various antibiotics such as chloramphenicol and tetracycline and to heavy metals such as arsenic and mercury |
0 |
WilkieLW Thu, 11 Mar 2010 19:39:49 GMT |
|
| F plasmids (fertility factors) |
direct the synthesis of proteins that self-assemble into conjugation pili. |
0 |
WilkieLW Thu, 11 Mar 2010 19:39:49 GMT |
|
Table 8.1
F' donor, F- recipient |
F' cell with some duplicate gene pairs: one on chromosome, one on plasmid. |
0 |
WilkieLW Thu, 11 Mar 2010 19:36:42 GMT |
|
Table 8.1
Hfr Donor, F- recipient |
F- with variable quantitiy of chromosomal DNA (F- recombinant) some of both? |
0 |
WilkieLW Thu, 11 Mar 2010 19:36:42 GMT |
|
Table 8.1
F+ Donor F- recipient |
F+ cells both Male |
0 |
WilkieLW Thu, 11 Mar 2010 19:36:42 GMT |
|
Conjugation
F+ cells and F- Cells |
F+ cells= male/donor cells
F- Cells= female/recipient cells
F+ cells contain extrachromosomal DNA called Fertility plasmids.
F+ cells make F pilus (sex pilus or conjugation pilus) a bridge by which it attaches to the F- cell.
F plasmids are circular, double stranded DNA molecules. |
0 |
WilkieLW Thu, 11 Mar 2010 19:36:42 GMT |
|
| Conjugation |
No cells die, both cells are alive.
Genetic info is transferred from one bacterial cell to another. Conjugation differs from those other mechanisms in two ways.
1. requires contact between donor and recipient cells.
2. transfers much larger quantities of DNA (occasionally whole chromosomes) |
0 |
WilkieLW Thu, 11 Mar 2010 19:17:14 GMT |
|
| Generalized Transduction |
bacteriophage infection of a host bacterium starts the lytic cycle. Bacterial chromosomes are fragmented and picked up to be packaged with the phage DNA into new phage particles. NO PARTICULAR AREA OF INTEGRATION. The new phages attack another host they carry the previous hosts DNA too. |
0 |
WilkieLW Thu, 11 Mar 2010 19:17:14 GMT |
|
| Specialized Transduction |
Several lysogenic phages are known to carry out specialized transduction. Lambda (h) phage in Escherichia coli has been extensively studied.
***phages usually insert at a SPECIFIC AREA when they integrate with a chromosome. (NEAR THE SITE OF PROPHAGE)Lambda phage inserts into the E. coli chromosome between the gal gene, which controls galactose use, and the bio gene, which controls biotin synthesis. |
0 |
WilkieLW Thu, 11 Mar 2010 19:17:14 GMT |
|
| lysogeny |
presistence of a prophage without phage replication and destruction of the bacterial cell. |
0 |
WilkieLW Thu, 11 Mar 2010 19:17:14 GMT |
|
| Prophage |
Combo of 2 DNA, DNA that is incorporated into the host bacterium's DNA. |
0 |
WilkieLW Thu, 11 Mar 2010 18:49:50 GMT |
|
| Temperate Phage |
ordinarily does not cause a distruptive infection. DNA is incorporated into a bacterium's DNA and replicated with it. |
0 |
WilkieLW Thu, 11 Mar 2010 18:49:50 GMT |
|
Bacteriophage Life Cycle
(Lytic Cycle) |
1. phage attaches to receptor site on bacterial cell wall, penetrates and inserts its DNA.
2. Phage DNA directs the bac cells metabolism to produce viral components- proteins and copies of phage DNA.
3. empty phage heads are synthesized.
4. heads are packed with DNA
5. collars, sheaths, and base plates attached to heads. Tail fibers added last.
6. Bacterial cell lyses (dies), releasing completed infective phages. |
0 |
WilkieLW Thu, 11 Mar 2010 18:49:50 GMT |
|
Bacteriophage Life Cycle
(Lysogenic Cycle) |
1. Phage attaches to receptor site on Bacteria. Releases DNA.
2. phage DNA inserts itself as a prophage into the bacterial chromosome.
3. phage is replicated along with Bacterial DNA
4. Binary fission complete each cell has the phage DNA incorporated.
(Can last for generations)
(Bacteria is not affected) |
0 |
WilkieLW Thu, 11 Mar 2010 18:49:50 GMT |
|
| Virulent Phage |
capable of causing infection and eventually, the distruction and death of a bacterial cell (Lytic Cell) |
0 |
WilkieLW Thu, 11 Mar 2010 18:35:09 GMT |
|
| Phages |
composed of a core of nucleic acid covered by a protein coat. Capable of infecting a bacterium attaches to a receptor site on the cell wall. Virus |
1 |
WilkieLW Thu, 11 Mar 2010 18:44:17 GMT |
|
| Bacteriophage |
a virus that can infect bacteria. |
0 |
WilkieLW Thu, 11 Mar 2010 18:35:09 GMT |
|
| Transduction |
is a method of transferring genetic material from one bacterium to another. Bacteriophages carry the DNA. |
0 |
WilkieLW Thu, 11 Mar 2010 18:35:09 GMT |
|
| Griffith's Experiment |
Discovery of Transformation=
Mice(pneumococcal)
S-type bacteria is alive and capable of killing.
R-type isn't capable of killing.
Heat killed S-type and R-type put in a mouse together produces transformation of DNA from the dead S-type picked up by the R-type and R-type is turned into Killer S-type. |
0 |
WilkieLW Thu, 11 Mar 2010 18:13:05 GMT |
|
| Naked DNA |
DNA that has been released from one org after the cell is lysed and DNA is no longer incorporated into chromosomes. S-type is lysed (dead) and R-type picks it up and becomes S-type. |
0 |
WilkieLW Thu, 11 Mar 2010 18:13:05 GMT |
|
| Transformation |
Competence factor=facilitiates the entry of DNA into a cell, DNA transport proteins and DNA exonuclease (an enzyme that cuts up DNA) is also needed. |
0 |
WilkieLW Thu, 11 Mar 2010 18:13:05 GMT |
|
| Transormation |
a change in an organism's characteristics because of the transfer of genetic information.
Dead bacteria leaves behind it's DNA, and another bacteria picks it up. |
0 |
WilkieLW Thu, 11 Mar 2010 18:13:05 GMT |
|
| Horizontal or Lateral gene transfer |
pass genes to other microbes of their same generation. |
0 |
WilkieLW Thu, 11 Mar 2010 18:01:50 GMT |
|
| Vertical gene transfer |
genes pass from parents to offspring (Bacteria) |
0 |
WilkieLW Thu, 11 Mar 2010 18:01:50 GMT |
|
| Recombination |
combining of genes (DNA) from two different cells. |
0 |
WilkieLW Thu, 11 Mar 2010 18:01:50 GMT |
|
| Gene Transfer |
movement of genetic information between organisms |
0 |
WilkieLW Thu, 11 Mar 2010 18:01:50 GMT |
|
