molecular bio 2 - applications Flashcards
A genome is all of a?
It is the entire set of ? building, running, and maintaining an organism, and passing life on to the next generation.” The whole shebang!!
*would mitochondria, plasmids and plastids be part of GENOME?
living things material
hereditary instructions for
heck yes!
Each species has its own distinctive genome: the dog genome, the human genome, the genomes of the cow, cold virus, bok choy, E. coli etc.
are mitochoondria unique to the cells of animals, plants and fungi?
main func of mitochondria?
their history?
DO THEY HAVE PROKARYOTIC OR EUKARYOTIC ORIGIN?
yes!
They have a double membrane, the inner layer usually folded inward to form
cristae
*metabolize or break down carbohydrates and fatty acids in order to generate energy (ATP)
- Symbiogenesis, or endosymbiotic theory, is an evolutionary theory: explains origin of eukaryotic cells from prokaryotes. It states that many imp. organelles of eukaryotes originated through symbiosis events.
(This theory was advanced and substantiated with microbiological evidence by Lynn Margulis in 1967)
- The endosymbiotic hypothesis for the origin of mitochondria suggests that mitochondria are descended from specialized bacteria, that somehow survived endocytosis by another cell type, and became incorporated into the cytoplasm.
*** MITOCHONDRIA AND OTHER ORGANELLES THEREFORE HAVE A PROKARYOTIC ORIGIN We can think of them as endosymbiotic bacteria which became cell organelles during evolution, and luckily remained!
* AS such, they have their own DNA (mtDNA or mDNA)
is plasmid big or small, is it straight or circular, single or double stranded? distinct from cell’s chromosomal DNA?
does it need a protein to replicate itself?
where do they exist?
Often, the genes carried in plasmids provide bacteria with genetic advantages, such as?
artificial plasmids are widely used as a vehicle for carrying a particular DNA (vector) for what?
** CAN PLASMIDS BE TRANSMITTED FROM ONE BACTERIA TO ANOTHER? AND WHAT IS IT CALLED?
A plasmid is a small, circular, double-stranded DNA molecule that is distinct from a cell’s chromosomal DNA, and can replicate independently. Plasmids naturally exist in bacterial cells, but can also occur in some Archaea and eukaryotes. Often, the genes carried in plasmids provide bacteria with genetic advantages, such as antibiotic resistance.
Artificial plasmids are widely used as vectors in molecular cloning, serving to drive the replication of recombinant DNA sequences within host organisms
- Plasmids can be transmitted from one bacterium to another
- This host-to-host transfer of genetic material is called horizontal gene transfer
What is a plastid? *** IMP they are sites of?
Apicoplast is derived from? found in most? apicoplast originated from “what” through “what”?
plastids are double membrane found in cells of plants and algae (chloroplast)
***and they are sites of manufacture and storage of imp, chemical compounds used by cell and are often used as drug targets for chemotherapy (e.g. malaria)
An apicoplast is a derived non-photosynthetic plastid found in most Apicomplexa, including malaria parasites such as Plasmodium falciparum, but not in others such as Cryptosporidium.
It originated from an alga (there is debate as to whether this was a green or red alga) through secondary endosymbiosis.
Horizontal gene transfer
*** IMP Serial endosymbiosis what is it?
- 1st was mitochondria (like Lynn Margulis was mentioning)
- and then plastid coming in in plants and then the evolution of algal cells occurring,
then red algal plastids being endosymbiosis second time and then remaining in the chromealveolate
serial endosymbiosis: Plastids (pigment-bearing organelles such as chloroplasts) evolved from a cyanobacterium that was engulfed by a heterotrophic eukaryote (primary endosymbiosis).
That ancestral eukaryote diversified into red algae and green algae, some of which were subsequently engulfed by other eukaryotes (secondary endosymbiosis); these plastids may retain their own nucleus, and contain genetic material
Techniques and applications
DNA hybridization and Southern blotting: what is it used tod detect?
Southern blotting combines transfer of electrophoresis-separated DNA fragments to a filter membrane and subsequent fragment detection by?
The northern blot is a technique used in molecular biology research to?
Western Blotting used to identify?
uses what to separate native proteins by 3-D structure or denatured proteins by the length of polypeptide?
which 2 a re similar in terms of use of gel electrophoresis?
certain dna SEQUENCES FROM DNA samples
probe hybridization.
northern blot to study gene expression by detection of RNA (or isolate mrNA) in a sample?
Western blotting (immunoblot) is a very useful technique used in molecular biology, to identify specific proteins from a complex mixture of proteins extracted from cells or an organ.
It uses gel electrophoresis to separate native proteins by 3-D structure or denatured proteins by the length of the polypeptide.
S and W
most common application of molecular biology?
What do they amplify?
*COMPONENTS OF it? IMP!!
Polymerase chain reaction (PCR) invented by Kary Mullis (oligonucleotide synthesis),
◼ Nobel Prize-winning American biochemist
◼ One of the monumental scientific techniques of
the twentieth century
Dr. Mullis received a Nobel Prize in chemistry in 1993
Amplify specific DNA fragments from genomic DNA, genes etc.
COMPONENTS of PCR rxn:
- Template DNA
- Oligonucleotide Primers (complementary single stranded DNA
- DNTP (deoxynucleotides- A, T, G, C)
- Taq polymerase (A very **thermostable polymerase) - makes it happen faster
- The correct conditions, e.g. buffers [Mg Cl2]
Remember the rRNA gene (rDNA) is a multiple copy gene, so there are multiple sites per DNA strand. Good for amplification; non-linear amplification or exponential!
PCR
process
NON-LINEAR AMPLIFICATION OR EXPONENTIAL
- denaaturation at 94-96 C (here H bonds separate so goes from 1 to 2 strands)
- Annealing 68 C
- Elongation 72 C
cycles repeated 30-35 times
Remember the rRNA gene (rDNA) is a multiple-copy gene, so there are multiple sites per DNA strand. Good for amplification; non-linear amplification or exponential!
