Download UNIT SIX: MOLECULAR GENETICS AND BIOTECHNOLOGY

Survey
yes no Was this document useful for you?
   Thank you for your participation!

* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project

Document related concepts

DNA repair wikipedia , lookup

Genome evolution wikipedia , lookup

Eukaryotic transcription wikipedia , lookup

RNA-Seq wikipedia , lookup

Agarose gel electrophoresis wikipedia , lookup

Genetic code wikipedia , lookup

Maurice Wilkins wikipedia , lookup

List of types of proteins wikipedia , lookup

Mutation wikipedia , lookup

Promoter (genetics) wikipedia , lookup

Gene expression wikipedia , lookup

Transcriptional regulation wikipedia , lookup

Gel electrophoresis of nucleic acids wikipedia , lookup

DNA vaccination wikipedia , lookup

Transformation (genetics) wikipedia , lookup

Gene wikipedia , lookup

Silencer (genetics) wikipedia , lookup

Community fingerprinting wikipedia , lookup

Molecular cloning wikipedia , lookup

Replisome wikipedia , lookup

DNA supercoil wikipedia , lookup

Vectors in gene therapy wikipedia , lookup

Biosynthesis wikipedia , lookup

Non-coding DNA wikipedia , lookup

Molecular evolution wikipedia , lookup

Cre-Lox recombination wikipedia , lookup

Nucleic acid analogue wikipedia , lookup

Artificial gene synthesis wikipedia , lookup

Deoxyribozyme wikipedia , lookup

Transcript
UNIT SIX: MOLECULAR GENETICS AND BIOTECHNOLOGY
MAIN IDEA: THE DISCOVERY THAT DNA IS THE GENETIC CODE INVOLVED MANY
EXPERIMENTS
OBJECTIVE 1: EXPLAIN THE IMPORTANCE OF DNA, ANALYZE ITS STRUCTURE, AND DESCRIBE
THE IMPORTANCE OF NUCLEOTIDE SEQUENCES
A. Genetic material held in the molecules of DNA ultimately determines an organism’s traits.
1. DNA achieves this control by making all the proteins for an organism.
a. Remember enzymes are proteins and these control the chemical reactions needed for life. .
B. Review structure of DNA
1. made of structural units called nucleotides
a. sugar – deoxyribose
b. Phosphate – one phosphorous atom surrounded by 4 oxygen atoms
c. Nitrogen base – a carbon ring that contains one or more atoms of nitrogen. There are four
bases: adenine (A), guanine (G), cytosine (C), and thymine (T), so DNA has four possible
nucleotides.
d. Nucleotides form long chains, with the phosphate group of one nucleotide bonding to the
deoxyribose sugar of the adjacent nucleotide.
e. The amount of adenine always equals the amount of thymine, and the amount of cytosine is
always equal to the amount of guanine: Chargoff’s rule (see page 329, Figure 12.5)
C. Four scientists joined the search for the DNA structure: Franklin, Wilkins, Crick and Watson
1. Rosalind Franklind’s Photo and x-ray diffraction data helped Watson and Crick solve the structure
of DNA. The photo indicated that DNA was a double helix a twisted ladder shape
2. Watson and Crick proposed the double helix structure of DNA (see page 331 figure 12.8):
a. Two outside strands of alternating deoxyribose and phosphate
b. Cytosine and guanine bases pair to each other by three hydrogen bonds.
c. Thymine and adenine bases pair to each other by two hydrogen bonds.
3. Two strands of DNA running antipparallel make up the DNA helix.
a. Carbon molecules can be numbered in organic molecules like sugar.
b. On the top the 5’ carbon is on the left and on the end of the rail the 3” is on the right. The
strand is oriented 5’ to 3”
c. The strand on the bottom is oriented 3’ to 5’.
4. In 1953, Watson and Crick published a one page letter suggesting the structure of DNA and
hypothesized a method for its replication. In the same issue, Franlkin and Wilkins presented
evidence supporting Watson and Crick.
5. The variation found among species is because of the varying sequence of the four different
nucleotides along the DNA strands.
OBJECTIVE 2: DESCRIBE THE BASIC STRUCTURE OF A EUKARYOTIC CHROMOSOME
A. (See Page 332, Figure 12.9) Eukaryote DNA is organized into chromosomes each having from 51
million to 245 million base pairs.
B. In order to fit into the eukaryotic cell, the DNA tightly coils around a group of beadlike proteins called
histones.
C. The phosphate groups in DNA create a negative charge, which attract the DNA to the positively
charged histone proteins and form a nucleosome. The nucleosomes then group together into
chromatin fibers, which supercoil to make up the DNA structure recognized as a chromosome.
MAIN IDEA: DNA REPLICATES BY MAKING A STRAND THAT IS COMPLIMENTARY TO EACH
ORIGINAL STRAND.
OBJECTIVE 3: DESCRIBE THE REPLICATION OF A DNA MOLECULE AND THE ROLE OF ENZYMES
IN THE REPLICATION PROCESS
A. Before a cell can divide by mitosis or meiosis, it must first make a copy of its chromosomes in
interphase. The DNA in the chromosomes is copied in a process called semiconservative
replication.
1. Parental strands of DNA separate, serve as templates, and produce DNA molecules that have
one strand of parental DNA and one strand of new DNA. (see pages 333 and 334)
2. Without DNA replication, new cells would have only half of the DNA of their parents; species
could not survive and individuals would not grow or reproduce successfully.
3. ALL organisms undergo DNA replication.
1
B. Many enzymes are needed to ensure proper DNA replication
1. DNA helicase is the enzyme responsible for unwinding and unzipping the double helix. This
enzyme helps break the hydrogen bonds that hold the two strands of DNA together.
2. Single stranded binding proteins associate with the DNA and keep the strands separate during
replication.
3. As the helix unwinds, another enzyme, RNA primase, adds a short segment of RNA called RNA
primer on each strand of DNA.
4. The enzyme DNA polymerase adds new nucleotides to the 3’ end of the new strand.
a. adenine bonds with thymine and cytosine bonds with guanine
C. Now the genetic make-up of an organism can be passed on to new cells during mitosis or to new
generations through meiosis followed by sexual reproduction.
OBJECTIVE 4: EXPLAIN HOW LEADING AND LAGGING STRANDS ARE SYNTHESIZED
DIFFERENTLY
A. When completing semiconservative replication, the two strands of DNA are made in a slightly
different manner.
B. One strand is called the leading strand and is elongated as the DNA unwinds.
1. It is built continuously with nucleotides added to the 3’ end.
C. The lagging strand elongates away from the replication fork.
1. It is synthesized discontinuously into small fragments called Okazaki fragments by the DNA
polymerase in the 3’ to 5’ direction.
2. An enzyme called DNA ligase will later connect these fragments.
D. So, not only is DNA replication called semiconservative but is also called semidiscontinuous.
E. Their may be many areas along the chromosome where replication began. When DNA polymerase
comes to an RNA primer on the DNA, it removes the primer and fills in the place with DNA
nucleotides. When the RNA polymerase has been replaced, DNA ligase links the two sections.
OBJECTIVE 5: COMPARE DNA REPLICATION IN EUKARYOTES AND PROKARYOTES
A. Eukaryotic DNA unwinds in multiple areas as DNA is replicated.
1. Each individual area of a chromosome replicates as a section, so multiple areas of replication are
occurring along the large eukaryotic chromosome at the same time. (see page 335, figure 12.12)
B. In prokaryotes, the circular DNA strand is opened at one origin of replication. (see page 335, figure
12.12)
1. Replication occurs in two directions just as in eukaryotic replication.
2. DNA replication is shorter than in eukaryotics and it remains in the cytoplasm, not packaged into
the nucleus.
MAIN IDEA: DNA CODES FOR RNA, WHICH GUIDES PROTEIN SYNTHESIS
OBJECTIVE 6: DESCRIBE HOW GENES ARE RELATED TO PROTEINS
A. Sequences of DNA contain information to produce proteins.
B. Review the structure and function of proteins. Enzymes (a type of protein) control all the chemical
reactions in an organism.
C. By encoding the instructions for making proteins, DNA controls cells.
D. The sequence of nucleotides in each gene contains information for assembling the string of amino
acids that make up a single protein. Each human cell contains about 80, 000 genes.
E. Geneticists now accept the basic mechanism for reading and expressing genes is from DNA to RNA
to protein.
OBJECTIVE 7: DESCRIBE THE STRUCTURE AND FUNCTION OF RNA
A. RNA (ribonucleic acid) is a nucleic acid and its structure is similar to that of DNA.
1. RNA is single stranded, with alternating chains of phosphate and sugar.
2. The sugar is ribose.
3. The nitrogen bases are adenine, guanine, cytosine, and uracil. Uracil pairs with adenine.
THERE IS NO THYMINE IN RNA!
B. RNA takes instruction from DNA and assembles the protein amino acid by amino acid.
C. There are three structural types of RNA that help to build proteins. (see page 336, table 12.2)
1. Messenger RNA (mRNA) molecules are long strands of RNA nucleotides that are formed
complimentary to one strand of DNA. They travel to the nucleus to the ribosome to direct the
synthesis of a specific protein.
2
2. Ribosomal RNA (rRNA) this type of RNA is associated with proteins to form ribosomes in the
cytoplasm
3. Transfer RNA (tRNA) smaller segments of RNA that transport amino acids to the ribosome.
OBJECTIVE 8: DESCRIBE THE PROCESSES OF TRANSCRIPTION AND TRANSLATION
A. Transcription: DNA à mRNA (see page 337)
1. DNA unzips, but only one side of the DNA acts as a template
2. RNA polymerase, an enzyme that regulates RNA synthesis, binds to a specific site where RNA
will be made.
3. As DNA unwinds, the RNA polymerase starts RNA synthesis along one strand of DNA in a 3’ to
5’ direction.
a. The strand that is read by RNA polymerase is called the template strand and RNA
nucleotides are added to the 3’ end
4. The complimentary nucleotide will bind temporarily to the DNA strand, acting as a template, so
that the information from the DNA is now transcribed to form mRNA
a. if the DNA base is G then the RNA base is C
b. if the DNA base is C then then RNA base is G
c. if the DNA base is T then the RNA base is A
d. if the DNA base is A then the RNA base is U
5. When the correct sequence of complimentary bases has been transcribed, the mRNA detaches
from the DNA
6. While in the nucleus, the RNA is modified. Introns (sequences of nucleotides that don’t carry
useful information for protein synthesis) are cut out and spliced together at exons, the coding
sequences that remain in the final mRNA. Once processed the mRNA carries the appropriate
codon sequence for protein and leaves the nucleus and enters the cytoplasm.
1. Other processing strategies of the pre-mRNA include adding a protective cap at the 5’ end
(thought to aid in ribosome recognition) and adding a tail of many adenine nucleotides called
a poly-A tail (function presently unknown)
7. The DNA strands zip back together.
8. Instructions for protein synthesis are encoded in DNA. The only way DNA varied among
organisms is in the sequence of nitrogenous bases.
1. Twenty amino acids are used to make proteins, so DNA must provide for at least 20 different
codes.
2. Based on math, it was hypothesized that a group of three bases coded for a single amino
acid. 64 combinations are possible when a sequence of three bases is used; thus 64
different mRNA codons are in the genetic code.
3. The code for protein synthesis, therefore is the order of the bases in a single strand of DNA.
Every three bases on mRNA (or every three nucleotides) are known as a codon and codes
for one amino acid. (see page 338 Fig. 12.4)
a. Some codons do not code for amino acids; they may be stop codons.
B. Translation: mRNA à tRNA à protein (see pages 339)
a. Once mRNA is synthesized and processed it moves out of the nucleus to the ribosome in the
cytoplasm.
b. The 5’ end of the mRNA attaches to rRNA (the ribosome), the workbench for protein
synthesis
c. There are many different types of tRNA and this molecule acts as as the interpreters of the
mRNA codon sequence.
1. Each tRNA carries a specific amino acid on its head.
2. Each tRNA has as series of three bases at its feet called an anticodon.
3. If a codon on the mRNA were CGA then the complimentary anticodon would be GCU.
4. The anticodon is read 3’ to 5’
d. The first complimentary tRNA lands on the mRNA and passes its amino acid to the second
tRNA
e. The third tRNA receives the two amino acids from the second tRNA.
f. A start codon signals the beginning of the sequence of codons to be translated and stop
codons end the sequence to be translated to a protein. Eventually a long chain of amino
acids is formed which is the protein.
g. Depending on the complexity of the organism, there may be 100 to several 1000 proteins as
part of its makeup.
3
h. Each of the 20 amino acids can appear many times in the protein molecule. The structure
and function of each protein depends on the sequence of amino acids.
i. All organisms use the same genetic code for amino acids and assembling proteins.
j. These proteins become enzymes and cell and tissue structures. The formation of protein,
originating from the DNA code, produces the diversity of our living world!
MAIN IDEA: GENE EXPRESSION IS REGULATED BY THE CELL, AND MUTATIONS AFFECT THIS
EXPRESSION
OBJECTIVE 9: DESCRIBE HOW BACTERIA (PROKARYOTES) ARE ABLE TO REGULATE THEIR
GENES BY TWO TYPES OF OPERONS
A. Gene regulation is the ability of an organism to control which genes are transcribed in response to
the environment.
B. In prokaryotes, an operon, often controls the transcription of genes in response to changes in the
environment.
1. An operon is a section of DNA that has genes for the proteins needed for a specific metabolic
pathway.
a. Parts of an operon are an operator, promoter, regulatory gene, and genes coding for proteins.
C. The operator is a piece of DNA that acts as an on/off switch for transcription.
nd
D. A 2 segment of DNA is called the promoter where the RNA polymerase first binds to DNA.
E. The trp operon is a repressible operon (see page 343, figure 12.7)
1. Synthesis of the amino acid tryptophan occurs in five steps and each step is catalyzed by an
enzyme.
2. The five genes coding for these enzymes are clustered together on the chromosome
3. Transcription of the five enzymes is normally repressed, or turned off. If tryptophan is in the
environment then the prokaryote doesn’t need to make it so the trp repressor gene turns off the
transcription process by making a repressor protein.
4. Tryptophan combines with the inactive repressor protein and this complex binds to the operator in
the promoter sequence. With the repressor bound to the operator, RNA polymerase cannot bind
to it, which prevents the transcription of the enzyme genes. This stops the synthesis of
tryptophan by the cell.
F. The lac operon is an example of the gene expression of inducible enzymes. (see page 343, figure
12.18) It is an inducible operon because transcription is turned on by an inducer.
1. Bacteria make an enzyme that let use lactose as an energy source.
2. The lactose operon contains a promoter, an operator, a regulatory gene, and three enzymes that
control lac digestion.
3. When the lac operon is “off”, the regulatory gene makes a repressor protein that binds to the
operator in the in the promoter sequence and prevents the transcription of the enzyme genes.
4. When the inducer is present, it binds to the repressor and inactivates it. The inducer is allactose,
a molecule that is present in foods that have lactose. SSSOOOO, when lactose is present, the
allactose binds to the repressor and inactivates it. With the repressor inactivated, RNA
polymerase can bind to the promoter and start transcription.
OBJECTIVE 10: DISCUSS HOW EUKARYOTIC REGULATE TRANSCRIPTION OF A GENE
A. In eukaryotic cells, many genes interact with one another, requiring more than one promoter and
operator for a set of genes.
B. One way that eukaryotes control gene expression is through proteins called transcription factors.
1. Transcription factors ensure that a gene is used at the right time and that proteins are made in the
right amount.
2. There are two main sets of transcription factors:
a. One factor forms complexes that guide and stabilize the binding of the RNA polymerase to
the a promoter.
b. Another set includes regulatory proteins that help control the rate of transcription.
C. The shape of eukaryotic DNA also regulates transcription. Because eukaryotic DNA is wrapped
around histones to form nucleosomes, this structure provides some inhibition of transcription.
1. Some regulatory proteins and RNA polymerase still can activate specific genes even if they are
packaged in the nucleosome.
D. Gene regulation is VERY important during development. As the zygote undergoes mitosis, it
produces all kinds of cells needed by the organism. During differentiation the cells will become
specialized.
4
1. Genes called Homeobox (hox) control differentiation.
2. Hox genes determine the body plan of an organism.
3. Hox genes code for transcription factors and are active zones of the embryo that are in the same
order as the genes on the chromosome. (see page 344, figure 12.19)
a. These genes, transcribed at specific times, and located in specific places on the genome,
control what body part will develop in a given location.
E. RNA interference (RNAi) is another way eukaryotes regulate genes.
1. An enzyme called dicer cuts small pieces of double stranded RNA. These small pieces are
called small interfering RNA.
2. The small interfering RNA binds to a protein complex that degrades one strand of RNA,
preventing its translation.
3. The use of RNAi as a genetic therapy to silence a specific gene that has gone bad has entered
human therapeutic clinical trials.
OBJECTIVE 11: DEFINE THE WORD MUTATION AND DIFFERENTIATE BETWEEN VARIOUS TYPES
A. Any change in the DNA sequence is called a mutation. The mutation may or may not effect the
expression of the gene or the sequence of amino acids in the encoded protein.
B. A point mutation is a change in a single base pair in DNA. A single change in a single nitrogen
base can change the entire structure of the protein because a change in a single amino acid can
affect the shape of the protein. It is enough to cause a genetic disorder. Types of point mutations
include the following:
1. Substitution – one base is exchanged for another
a. missense substitutions code for the wrong amino acid
b. nonsense substitutions change the codon for an amino acid to a stop codon. These cause
translation to stop early. Nearly all of these make proteins that are nonfunctional.
c. The change of one amino acid for another amino acid in a protein can affect the folding and
stability of the protein. (see page 347, figure 12.22) This is the cause of sickle cell disease, in
which the structure of hemoglobin is changed.
C. A frameshift mutation occurs when a single base is added or deleted from DNA causing a change
in the chromosome structure.
1. Insertions – gain of a nucleotide
2. Deletion – loss of a nucleotide
3. This would cause nearly every amino acid in the protein after the deletion or addition to be
changed.
D. In general, point mutations are less harmful to an organism than a frameshift mutation because they
disrupt only a single codon.
E. Sometimes mutations are associated with diseases and disorders. (See page 346, Table 12.3)
F. Large portions of DNA can also be involved in a mutation and have drastic effects on the expression
of these genes.
1. A piece of an individual chromosome containing one or more genes can be deleted or moved to a
different location on the chromosome or even to a different chromosome
a. Inversions – occur when part of a chromosome breaks off and is reinserted backwards.
b. Translocations – occur when part of one chromosome breaks off and is added to a different
(non-homologous) chromosome.
G. In 1991, a new kind of mutation was discovered that involves an increase in the number of copies
repeated codons, called tandem repeats.
1. Fragile X syndrome is due to many repeated CGG units near the end of the X chromosome,
making the lower tip of the X chromosome appear fragile. (see page 347, Figure 12.21)
H. Some mutations result from a change in the number of chromosomes (review unit 4)
1. Changes in numbers result in monosomy, trisomy, and polyploidy.
2. Few chromosomes mutations are passed on to the next generation because the zygote usually
dies. In cases where the zygote survives, the mature organism is usually sterile and incapable of
producing offspring.
3. Nondisjunction is a type of chromosome mutation that occurs frequently in plants and can be
beneficial.
5
OBJECTIVE 12: DESCRIBE CAUSES OF MUTATIONS AND EXPLAIN HOW DNA CAN REPAIR
ITSELF
A. Some mutations are spontaneous…they just happen, especially point mutations
1. During replication, DNA polymerase may add the wrong nucleotides, but because DNA
polymerase has a proofreading mechanism, the wrong nucleotide gets added only for one in
100,000 bases; it goes unfixed in less than 1 in 1,000,000,000.
2. The greater the exposure to the mutagen, the more likely the chance that a mistake will not be
corrected.
B. Many mutations are caused by factors in the environment.
1. Any agent that can cause a change in DNA is called a mutagen.
2. Mutagens include high-energy radiation, chemicals, and even high temperatures.
C. Many chemicals are mutagens and have a variety of affects. Chemical mutagens may be found in the
environment or in building materials: dioxin, asbestos, benzene, cyanide, and formaldehyde.
1. Some chemical mutagens have chemical structures that look like nucleotides so closely that they
substitute for them. If they become part of DNA it can replicate properly.
a. Medically this type of chemical has become useful: many drugs used to treat HIV mimic
various nucleotides. Once incorporated into the viral DNA, the DNA cannot copy itself.
2. Some affect DNA by changing the chemical structure of the bases. Bases may misrepair or bond
with wrong bases.
D. Forms of radiation, x-rays, cosmic rays, and nuclear radiation are highly mutagenic.
1. When radiation reaches the DNA, electrons absorb energy and may escape from their atom,
leaving behind what is called a free radical. Free radicals are charged atoms with unpaired
electrons that react violently with other molecules, including DNA.
2. U.V. radiation does not create free radicals but can cause adjacent thymine bases to bind to one
another, causing a disruption or “kink” in the DNA and preventing the replication of the DNA.
OBJECTIVE 13: DIFFERENTIATE BETWEEN A BODY CELL AND A SEX CELL MUTATION
A. If the mutation in a body cell (somatic cell) escapes the repair mechanism, it can be passed on to the
daughter cell. This does not affect future generations.
1. Some mutations might be neutral and might not cause problems to the cell if:
a. the mutation is in a sequence that is not used by the adult cell
b. the mutation occurred in an exon
c. the mutation may have not changed the amino acid coded for.
2. Some mutations may result in the production of an abnormal protein, causing inability for cell
function or even cell death.
3. If the mutation affects the cell cycle, the result can be cancer.
B. When mutations occur in the sex cells, also called sperm line cells, the mutations are passed on to
the organism’s offspring.
1. These mutations might not affect the function of the cell in the organism, but DRAMATICALLY
affect the offspring.
MAIN IDEA; RESEARCHERS USE GENETIC ENGINEERING TO MANIPULATE DNA
OBJECTIVE 14: EXPLAIN THE CONCEPT OF GENETIC ENGINEERING AND DISCUSS THE TOOLS
USED TO ENGINEER TRANSGENIC ORGANISMS
A. Genetic engineering involves cutting DNA from one organism into small fragments and inserting the
fragments into a host organism of the same or different species.
1. Also called recombinant DNA technology (connecting or recombining DNA fragments from
different sources).
2. Recombinant DNA technology involves the recombination of specific pieces of DNA, in the lab,
between pre-selected organisms to achieve a specific goal. The scientist, rather than natural
selection, then determines the usefulness of the recombinant DNA.
3. Can be used to quickly and reliably increase the frequency of a specific allele in a population,
much faster than selective breeding.
4. Genetically engineered organisms are used to study the expression of a specific gene,
investigate cellular processes, study the development of a certain disease, and select traits that
might be beneficial to people.
B. An organism’s genome is the total DNA present in the nucleus of each cell. Genomes can contain
millions and millions of nucleotides in the DNA within genes.
6
C. DNA tools can be used to manipulate DNA and to isolate genes from the rest of the genome.
1. Fragments of DNA are cut (digested) from a larger chromosome using restriction enzymes –
bacterial proteins that can cut both strands of the DNA molecule at specific nucleotide
sequences.
a. Restriction enzymes are also called endonucleases and are used by bacteria for defense
against viruses.
b. Eco RI is is a restriction enzyme used by scientists to cut DNA at a specific site – GAATTC.
c. Now the double stranded DNA has single-stranded ends which attract other single stranded
ends. The ends are “sticky”.
d. If the same enzyme is used to cut DNA from two organisms, the two pieces of DNA will have
matching sticky ends and will join or recombine. (see page 364, Figure 13.4)
2. Gel electrophoresis is used to separate DNA fragments according to their size by using an
electric current. (see page 365, Figure 13.5)
a. Form little gel squares with holes (wells) in one end
b. Place some of the cut pieces of DNA in one of the wells that are negatively charged
c. Submerge the gel under water
d. Add electricity to the water
e. The DNA fragments move to the positive end of the gel, sorting themselves according to size.
Larger pieces with more base sequences move the least distance.
f. The unique pattern created based on size of the DNA fragment can be compared to known
DNA fragments for identification.
OBJECTIVE 15: EXPLAIN HOW RECOMBINANT DNA IS MADE AND THEN USED IN GENE
CLONING, DNA SEQUENCING, AND POLYMERASE CHAIN REACTION
A. When DNA fragments have been separated by gel electrophoresis, fragments of a specific size can
be removed from the gel and combined with DNA fragments from another source. This newly
generated DNA molecule with DNA from different sources is called recombinant DNA. This enables
individual genes to be studied.
B. Many steps are used in forming recombinant DNA and large quantities of molecules are needed in
order to study it.
1. First, carriers called vectors transfer the recombinant DNA into the bacterial cell called the host
cell. Vectors can be biological (viruses or rings of DNA from bacteria called plasmids ) or they
can be mechanical. Host cells can also be a plant or animal.
a. Plasmids (a ring of DNA) are useful because they can be cut with restriction enzymes.
2. Gene splicing or ligation rejoins the DNA fragments. (see page 366, Figure 13.6)
a. If a plasmid and a DNA fragment from another genome have been cleaved by the same
restriction enzyme, their ends will be complimentary and can be combined.
b. An enzyme normally used by cells in DNA repair and replication, called DNA ligase, joins the
two DNA fragments chemically.
C. The recombinant DNA can now be inserted into s host cell so that lots of the recombinant DNA can
be made.
D. Large numbers of identical bacteria, each containing the inserted recombinant DNA, can be made in
a process called cloning.
1. To make a large quantity of recombinant plasmid DNA, bacterial cells are mixed with recombinant
DNA. Some bacterial cells take up the recombinant plasmid DNA through a process called
transformation. (See page 367, Figure 13.7)
a. When heated temporary openings in the plasma membrane allow recombinant DNA plasmid
to enter the bacterial cell. The bacterial cells make copies of the recombinant plasmid DNA
during replication.
E. DNA can be sequenced to provide scientists with much information:
1. Used to predict the gene’s function
2. Enables scientists to compare the genes with similar sequences from other organisms
3. Enables scientists to identify mutations or errors in the DNA sequence
a. before sequencing the DNA must be digested using restriction enzymes
4. See page 368 to understand how DNA is sequenced
F. Polymerase chain reaction (PCR) can be used to make millions of copies of a specific region of a
DNA fragment, once the DNA has been sequenced.
1. PCR is used for research in laboratories, by forensic scientists to identify suspects and victims in
criminal investigations, and by doctors studying and detecting infectious diseases.
G. See page 370, Table 13.1 to review tools/ process, function and application of genetic engineering.
7
OBJECTIVE 16: DEFINE BIOTECHNOLOGY AND DESCRIBE HOW TRANSGENIC ORGANISMS
MAY BE BENEFICIAL
A. Biotechnology, the use of genetic engineering to find solutions to problems, makes it possible tpp
produce organisms that contain individual genes from other organisms.
1. Organisms that have been genetically engineered by inserting genes from another organism are
called transgenic organisms.
B. Transgenic organisms are used in research and include bacteria, plants and animals.
1. Transgenic animals
a. most are made in the lab for research, often to study diseases and how to treat them. Mice,
fruit flies and roundworms are commonly used.
b. Livestock have been produced to improve the food supply and human health. Transgenic
goats have been engineered to secrete a protein, antithrombin III, which is used to prevent
human blood from clotting during surgery.
c. Transgenic fish have been produced to grow faster.
2. Transgenic plants
a. Many species have been developed to be resistant to insect and viral pests.
b. Researchers are developing peanuts and soybeans that don’t cause allergic reactions.
c. Many types are foods that would improve the human condition:
(1) sweet potato plants that would withstand a virus that typically wipes out most of the
African harvest
(2) rice with increased iron and vitamins that could decrease malnutrition in Asia
(3) bananas that produce vaccines for infectious diseases
(4) plants that produce biodegradable plastics
C. Transgenic bacteria
1. Insulin growth hormones, and substances that dissolve blood clots are made by transgenic
bacteria.
2. Other transgenic bacteria prevent frost damage on crops, clean up oil spills, and decompose
garbage.
MAIN IDEA: GENOMES CONTAINALL OF THE INFORMATIONNEEDED FOR AN ORGANISM TO
GROW AND SURVIVE
OBJECTIVE 17: DESCRIBE COMPONENTS OF THE HUMAN GENOME
A. A genome is the complete genetic information of a cell. The Human Genome Project (HGP) was an
international project that was completed in 2003 and its goal was to
1. determine the sequence of the approximately 3 billion nucleotides that make up human DNA
2. identify all of the 20,000 to 50,000 human genes.
B. Scientists have studied the genome of other organisms as a way to help develop the technology
needed to handle the large amounts of data generated by the HGP. Technologies help interpret the
function of newly identified genes.
C. In order to determine one continuous human genome sequence, each of the 46 human
chromosomes was cleaved.
1. Restriction enzymes cut the DNA with overlapping sequences, then the fragments are combined
with vectors making recombinant DNA, which is then cloned and sequenced using an automated
sequencing machine.
2. DNA sequencing can be used to identify defective genes by sequencing the DNA of people with a
specific disease and comparing the sequence who do not have the disease.
D. After sequencing the genome, scientists found that less than 2% of all the nucleotides in the human
genome code for all the proteins in the body.
1. The genome must be filled with long stretches of repeated sequences that have no direct function.
These are called noncoding sequences.
OBJECTIVE 18: DESCRIBE HOW FORENSIC SCIENTISTS USE DNA FINGERPRINTING
A. The long, noncoding sequences of DNA are unique to each individual. So when these regions are
cut by restriction enzymes, the set of DNA fragments that are made are unique to the individual too.
B. DNA fingerprinting involves separating these DNA fragments using gel electrophoresis in order to
observe the distinct banding patterns that are unique to each person.
C. DNA fingerprinting can be used to identify suspects and victims in criminal cases, determine
paternity, and identify soldiers killed in war.
8
1. Genetic information can be obtained from blood, hair, semen and skin.
2. PCR is used to copy small amounts of DNA. DNA is then digested using restriction enzymes.
DNA fragments are then separated by gel electrophoresis and compared to DNA fragments of known
sources such as victims an suspects.
D. Since its development in 1985, DNA fingerprinting has also been used to exonerate those wrongfully
convicted
E. Pharmacogenomics
1. Pharmacogenomics is the study of how genetic inheritance affects the body’s response to
drugs. Benefits include more accurate dosing of drugs that are safer and more specific.
2. Might allow for drugs to be custom-made for a specific individual based on their genetic makeup.
3. Prescribing drugs based on someone’s individual makeup will increase safety, speed recovery
and decrease side effects.
F. Gene Therapy
1. This is a technique aimed at correcting mutated genes that cause human diseases.
2. Scientists insert a normal gene into a chromosome to take the place of a dysfunctional gene.
3. In most cases, a normal gene is fused with a viral vector to create recombinant DNA. Target cells
in the patient are infected with the virus and the recombinant DNA material is released into the
affected cells.
4. Once in the normal cells the gene inserts itself into the patients genome and starts functioning.
5. In 2003 the FDA stopped gene therapy trials in the U.S. after the death of a patient caused by a
reaction to the viral vector.
9