Ch 20 DNA Tools and Biotechnology

__ refers to the manipulation of organisms (living things) or their components to make useful products

Biotechnology

Making Multiple Copies of a Gene or Other DNA Segment To work directly with specific genes, scientists prepare well-defined DNA segments in multiple identical copies by a process called __ (The production of multiple copies of a specific DNA segment)

DNA cloning

Studying the Expression of Interacting Groups of Genes __ compare patterns of gene expression in different tissues at different times or under different conditions

DNA microarray assays

The main technologies for sequencing and manipulating DNA are called ___

DNA technology

Making Multiple Copies of a Gene or Other DNA Segment ___ is useful for two basic purposes: to make many copies of, or amplify, a particular gene and to produce a protein product from the gene for research, medical, or other purposes

Gene cloning

Human Gene Therapy and Gene Editing __ is the introduction of genes into an afflicted individual for therapeutic purposes and may be useful for treating disorders traceable to a single defective gene

Gene therapy

20.1: DNA sequencing and DNA cloning are valuable tools for genetic engineering and biological inquiry __is the direct manipulation of genes for practical purposes

Genetic engineering

Reproductive Cloning of Mammals Another example of nonidentity in clones is the first cloned cat named__ , who was the first cloned cat; however, CC differed somewhat from her female "parent"

CC (for carbon copy)

Human Gene Therapy and Gene Editing In 2014, a group of researchers reported using __ to correct a genetic defect in mice

CRISPR-Cas 9

Editing Genes and Genomes The ___ system is a powerful new technique for gene editing in living cells and organisms

CRISPR-Cas9

Reproductive Cloning of Mammals In 1997, Scottish researchers announced the birth of Dolly, a lamb cloned from an adult sheep by nuclear transplantation from a differentiated __

mammary cell

Cloning Plants: Single-Cell Cultures In plants, ___ can dedifferentiate and then give rise to all the specialized cell types of the organism A __ cell, such as this, is one that can generate a complete new organism (Differentiated cells taken from the root (the carrot) and incubated in culture medium could grow into normal adult plants, each genetically identical to the parent plant.)

mature cells totipotent

Studying the Expression of Interacting Groups of Genes A ____ consists of tiny amounts of many single-stranded DNA fragments representing different genes fixed to a glass slide in a tightly spaced array, or grid, of dots. (available for base pairing) Ideally, these fragments represent all the genes of an organism.

microarray

Environmental Cleanup Some modified __ can be used to extract minerals from the environment or degrade potentially toxic waste materials

microorganisms

Editing Genes and Genomes Researchers have also modified the technique so a gene with a __ in it can be repaired

mutation

DNA Sequencing A gene's complete nucleotide sequence can be determined using a process called ___ (the process of determining the sequence of nucleotides (As, Ts, Cs, and Gs) in a piece of DNA) The first automated procedure used a technique called ___ or ____ developed by Frederick Sanger. In this technique, one strand of a DNA fragment is used as a template for synthesis of a nested set of complementary fragments; these are further analyzed to yield the sequence.

DNA sequencing dideoxy or chain termination sequencing

Cloning Animals: Nuclear Transplantation Differentiated cells from animals generally do not divide in culture, much less develop into the multiple cell types of a new organism. Therefore, early researchers had to use a different approach to answer the question of whether differentiated animal cells are totipotent. Their approach was to remove the nucleus of an unfertilized egg cell or zygote and replace it with the nucleus of a differentiated cell, a procedure called ___

nuclear transplantation

Embryonic and Adult Stem Cells Embryonic stem cells are ___, capable of differentiating into many different cell types

pluripotent

Diagnosis and Treatment of Diseases Scientists can diagnose many human genetic disorders by using __ and sequence-specific__, then sequencing the amplified product to look 4 the disease-causing mutation.

PCR and primers

Making Multiple Copies of a Gene or Other DNA Segment __ are small, circular DNA molecules that replicate separately from the bacterial chromosome.

Plasmids

Other Methods for Studying Gene Function Another method for silencing expression of specific genes doesn't alter the genome; instead, it exploits the phenomenon of ___ In this experimental approach, double stranded RNA molecules that match the sequence of a particular gene are processed into a siRNAs that either block translation or trigger the degradation of the the gene's messenger RNA.

RNA interference (RNAi)

Medical Applications By using __ and ___ or other techniques to compare gene expression in healthy and diseased tissues, researchers are finding genes that are turned on or off in particular diseases. These genes and their products are potential targets for prevention or therapy.

RNA sequencing and microarray assays

Other Methods for Studying Gene Function ___, single nucleotide variants, are among the most useful genetic markers

SNPs (single nucleotide polymorphisms) This diagram depicts the same region of the genome from two groups of individuals, one group having a particular disease or condition with a genetic basis. Unaffected people have an A/T pair at a given SNP locus, while affected people have a C/G pair there. Once the allele is confirmed as being associated with the disease in question, the SNP that varies in this way can be used as a marker for the disease-associated allele.

Stem Cells of Animals __ are relatively unspecialized cells that can both reproduce indefinitely and under certain conditions, differentiate into 1 or more specialized cell types Thus, stem cells are able to both replenish their own population and generate cells that travel down specific differentiation pathways.

Stem cells

Amplifying DNA: The Polymerase Chain Reaction (PCR) and Its Use in DNA Cloning The key to PCR is an unusual, heat-stable DNA polymerase called ___ (optimal temperature= 72°C)

Taq polymerase

Human Gene Therapy and Gene Editing __ avoids the complications of using viral vectors to deliver gene therapy (Viral vectors= viruses that are used to carry genes into the body, they could infect healthy cells as well as cancer cells)

The CRISPR-Cas 9 system

Protein Production by "Pharm" Animals __ are pharmaceutical "factories," producers of large amounts of otherwise rare substances for medical use

Transgenic animals This transgenic goat carries a gene for a human blood protein, antithrombin, which she secretes in her milk. Patients with a rare hereditary disorder in which this protein is lacking suffer from formation of blood clots in their blood vessels. Easily purified from the goat's milk, the protein is used to prevent blood clots in these patients during surgery or childbirth.

Agricultural Applications DNA technology is being used to improve ___ productivity

agricultural

Synthesis of Small Molecules for Use as Drugs Using drugs that target particular molecules is feasible for treatment of ___ in which the molecular basis is well understood

cancers

Reproductive Cloning of Mammals Since 1997, ___ has been demonstrated in many mammals, including mice, cats, cows, horses, mules, pigs, and dogs

cloning

Making Multiple Copies of a Gene or Other DNA Segment In genetic engineering, a plasmid acts as a ___ , a DNA molecule that can carry foreign DNA into a host cell and be replicated there

cloning vector

Editing Genes and Genomes Cas-9 will cut both strands of any DNA sequence ___ to the guide RNA

complementary

Agricultural Applications Agricultural scientists have endowed (establish) a number of ___ with genes fro desirable traits such as delayed ripening and resistance to spoilage, disease, and drought.

crop plants

Embryonic and Adult Stem Cells Many __ contain stem cells capable of giving rise to differentiated embryonic cells of any type. Stem cells can be isolated from early embryos at a stage called the blastula stage or its human equivalent, the blastocyst stage. In culture, these __ cells reproduce indefinitely, and depending on culture conditions, can be made to differentiate into a variety of specialized cells

early embryos embryonic stem (ES)

Forensic Evidence and Genetic Profiles The probability that two people who are not identical twins have the same STR markers (2-5 DNA nucleotide sequence) is __

exceptionally small

Using Restriction Enzymes to Make a Recombinant DNA Plasmid To check the recombinant plasmids after they have been copied many times in host cells, a researcher might cut the products again using the same restriction enzyme, expecting two DNA fragments: one the size of the plasmid and one the size of the inserted DNA. To separate and visualize the fragments, researchers carry out a technique called ___ This technique uses a gel made of ___ to separate a mixture of nucleic acids or proteins based on size, charge, or other physical properties

gel electrophoresis a polymer

Amplifying DNA: The Polymerase Chain Reaction (PCR) and Its Use in DNA Cloning After a __ has been cloned, its protein product can be produced in larger amounts for research or practical applications.

gene

Making Multiple Copies of a Gene or Other DNA Segment This production of multiple copies of a single gene is a type of DNA cloning called __

gene cloning

Medical Applications One benefit of DNA technology is identification of human genes in which mutation plays a role in ___ These discoveries may lead to ways of diagnosing, treating, and even preventing such conditions.

genetic diseases

Forensic Evidence and Genetic Profiles An individual's unique set of genetic markers, or __, can be obtained by analysis of tissue or body fluids (A genetic marker is a gene or DNA sequence with a known location on a chromosome that can be used to identify individuals or species.)

genetic profile

Amplifying DNA: The Polymerase Chain Reaction (PCR) and Its Use in DNA Cloning A 3-step cycle- __, __, __,- brings about a chain reaction that produces an exponentially growing population of identical DNA molecules

heating, cooling, and replication

Agricultural Applications Genetic engineering in plants has been used to transfer many useful genes, including those for __ or __, increased resistance to salinity, and improved nutritional value of crops

herbicide, pest resistance

Editing Genes and Genomes Molecular biologists have long sought techniques for altering, or editing, the genetic material of cells or organisms in a predictable way. In one such technique, called ____ , specific mutations are introduced into a cloned gene, and the mutated gene is returned to a cell in such a way that it disables ("knocks out") the normal cellular copies of the same gene. If the introduced mutations alter or destroy the function of the gene product, the phenotype of the mutant cell may help reveal the function of the missing normal protein.

in vitro mtuagenesis

Induced Pluripotent Stem (iPS) Cells Researchers transformed the differentiated cells into a type of ES cell by using a retrovirus to introduce extra, cloned copies of four "stem cell" master regulatory genes. The "deprogrammed" cells are known as ___ cells because, in using this fairly simple laboratory technique to return them to their undifferentiated state, pluripotency has been restored. The four genes induced differentiated skin cells to become pluripotent stem cells, with characteristics of embryonic stem cells.

induced pluripotent stem (IPS)

DNA Sequencing More and more often, next-generation sequencing is complemented (or in some cases replaced) by ___, with each new technique being faster and less expensive than the previous one. (developed now)

"third-generation sequencing"

DNA Sequencing In the last 15 years, ____ techniques have been developed that are much faster

"next-generation sequencing"

Diagnosis and Treatment of Diseases Many envision a future of ___ where each person's genetic profile can inform them about diseases and conditions for which they may be at risk

"personalized medicine"

Protein Production in Cell Cultures __ and __ are among the first such products made in this way

Human insulin and human growth hormone (HGH)

Forensic Evidence and Genetic Profiles __ is used to amplify particular short tandem repeats (STRs), using sets of primers that are labeled with different-colored fluorescent tags; the length of the region, and thus the number of repeats, can then be determined by ___

PCR gel electrophoresis

Diagnosis and Treatment of Diseases __ may be associated with a disease-causing mutation

SNPs

Amplifying DNA: The Polymerase Chain Reaction (PCR) and Its Use in DNA Cloning The ___, can produce many copies of a specific target segment of DNA

polymerase chain reaction, PCR

Human Gene Therapy and Gene Editing __ can be used to deliver the normal genes into somatic cells, which should be able to multiply through the patient's life, for full effectiveness

viruses

Reproductive Cloning of Mammals Dolly's __ in 2003, as well as that of another cloned sheep from another experiment, led to speculation that this sheep's cells were not as healthy as those of a normal sheep

premature death

Amplifying DNA: The Polymerase Chain Reaction (PCR) and Its Use in DNA Cloning PCR uses a pair of ___ specific for the sequence to be amplified

primers

Making Multiple Copies of a Gene or Other DNA Segment To clone pieces of DNA using bacteria, researchers first obtain a plasmid (originally isolated from a bacterial cell and genetically engineered for efficient cloning) and insert DNA from another source ("foreign" DNA) into the plasmid. The resulting plasmid is now a ___ , a molecule containing DNA from two different sources, very often different species.

recombinant DNA molecule

Using Restriction Enzymes to Make a Recombinant DNA Plasmid A restriction enzyme usually makes many cuts, yielding ___

restriction fragments

Using Restriction Enzymes to Make a Recombinant DNA Plasmid Bacterial restriction enzymes cut DNA molecules at specific DNA sequences called __

restriction sites

Forensic Evidence and Genetic Profiles Genetic profiles are currently analyzed using genetic markers called ___ (These are tandemly repeated units of two- to five-nucleotide sequences in specific regions of the genome. The number of repeats present in these regions is highly variable from person to person (polymorphic), and even for a single individual, the two alleles of an STR may differ from each other.)

short tandem repeats (STRs)

Concept 20.3: Cloned organisms and stem cells are useful for basic research and other applications A __ is relatively unspecialized cell that can reproduce itself indefinitely, or under certain conditions can differentiate into 1 or more types of specialized cells

stem cell

Using Restriction Enzymes to Make a Recombinant DNA Plasmid The most useful restriction enzymes cleave the sugar-phosphate backbones in the two DNA strands in a staggered manner, the resulting double-stranded restriction fragments have at least one single-stranded end, called a ___ These short extensions can form hydrogen-bonded base pairs with complementary sticky ends on any other DNA molecules cut with the same enzyme. The associations formed in this way are only temporary but can be made permanent by ____, an enzyme which catalyzes the formation of covalent bonds that close up the sugar-phosphate backbones of DNA strands

sticky end (sticky ends join together because of complementary base pairing) DNA ligase

Protein Production by "Pharm" Animals __ animals are made by introducing genes from one species into the genome of another animal

transgenic