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Chapter 20 Reading Quiz. Genes from two different sources that are combined result in ____. Where are “sticky ends” found? What structures, naturally found in bacteria, cut up foreign DNA? What is the point of the genetic technology “PCR”?

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Chapter 20 Reading Quiz

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Chapter 20 Reading Quiz

  • Genes from two different sources that are combined result in ____.

  • Where are “sticky ends” found?

  • What structures, naturally found in bacteria, cut up foreign DNA?

  • What is the point of the genetic technology “PCR”?

  • Organisms whose genomes carry genes from another species are known as _____.

1. Explain how advances in recombinant DNA technology have helped scientists study the eukaryotic genome.

  • It resulted in the appearance of the biotechnology industry

  • We can now move genes across the species barrier

  • The Human Genome Project  transcribe and translate the entire human genome

  • We hope to improve health and food production 

2. Describe the natural function of restriction enzymes.

  • They are major tools in recombinant DNA technology

  • They occur naturally in bacteria where they protect the bacterium against intruding DNA

  • This protection involves restriction in which foreign DNA is cut into small segments 

3. Describe how restriction enzymes and gel electrophoresis are used to isolate DNA fragments.

  • Restriction enzymes only recognize short, specific nucleotide sequences called recognition sequences or restriction sites

  • Gel electrophoresis is used to separate either nucleic acids or proteins based upon molecular size, charge, and other physical properties

     characteristic banding 

4. Explain how the creation of sticky ends by restriction enzymes is useful in producing a recombinant DNA molecule.

  • They are used in the laboratory to join pieces of DNA from different sources (sometimes cells or even different organisms)

  • These are temporary unions  only held by a few hydrogen bonds 

5. Outline the procedures for producing plasmid and phage vectors.

  • Isolation of vector and gene-source DNA

    - bacterial plasmids & foreign DNA with gene of interest

    2. Insertion of gene-source DNA into the vector

    - plasmid DNA cut  mix foreign DNA and plasmids  add DNA ligase to catalyze new covalent bonds

  • Introduction of cloning vector into bacterial cells

    - naked DNA added to bacterial culture, plasmid DNA taken up by transformation

  • Cloning of cells (and foreign DNA)

    - reproduced and cloned

    5. Identification of cell clones carrying the gene of interest (indicators added) 

6. Explain how vectors are used in recombinant DNA technology.

  • Cloning vector  a DNA molecule that can carry foreign DNA into a cell and replicate there

  • Most often used: bacterial plasmids and viruses 

7. List and describe the two major sources of genes for cloning.

  • DNA isolated directly from an organism

    2. Complementary DNA made in the lab from mRNA templates 

8. Describe the function of reverse transcriptase in retroviruses and explain how they are useful in recombinant DNA technology.

  • It is the enzyme that transcribes DNA from an RNA template

    Viral genomic RNA

    (reverse transcriptase)

    Viral DNA

  • Useful in that we can utilize it’s natural function to incorporate genes of interest 

9. Describe how bacteria can be induced to produce eukaryotic gene products.

  • Expression vectors allow the synthesis of many eukaryotic proteins in bacterial cells

  • Expression vectors contain a prokaryotic promoter just upstream of a restriction site where the eukaryotic gene can be inserted

  • The bacterial host cell recognizes the promoter and proceeds to express the foreign gene that has been linked to it 

10. List some advantages for using yeast in the production of gene products.

  • Yeast cells are easy to grow as bacteria and they contain plasmids

  • Some recombinant plasmids combine yeast and bacterial DNA and can replicate in either

  • Posttranslational modifications required to produce eukaryotic proteins can occur 

11. List and describe four complementary approaches used to map the human genome.

  • Genetic mapping (linkage)  construct a linkage map with several thousand genetic markers

  • Physical mapping  ordering the DNA fragments

    - a map made by cutting the DNA of each chromosome into a number of identifiable fragments

  • Sequencing DNA  the complete nucleotide sequence of a genome is the ultimate map

  • PCR amplification  amplify DNA in amounts sufficient for study (polymerase chain reaction)

  • Chromosome walking  start with a known gene and “walk” along the DNA from that locus 

12. Explain how RFLP analysis and PCR can be applied to the human genome project.

  • RFLP (restriction fragment length polymorphisms)  differences in restriction fragment length that reflects variations in homologous DNA sequences

    - used as genetic markers for making linkage maps

    - are a “genetic fingerprint”

  • PCR (polymerase chain reaction)  allow any piece of DNA to be quickly amplified (copied) in vitro

    - produces linkage maps without the need for large family pedigree analysis 

13. Describe how recombinant DNA technology can have medical applications such as diagnosis of genetic disease, development of gene therapy, vaccine production, and development of pharmaceutical products.

  • Disease  DNA technology to diagnose, early detection, and identification of carriers

  • Gene therapy  traceable genetic disorders may eventually be correctable = normal genes introduced

    - many social and ethical questions

  • Vaccines  produce specific protein molecules which could cause immune response; gene splicing could make vaccine pathogens safer

  • Pharmaceutical products  make mostly proteins

    - gene splicing applies to insulin and growth hormone (genetically engineered human insulin) 

14. Describe how gene manipulation has practical applications for agriculture.

  • Essentially to improve productivity

    Animals  bovine growth hormone (made by ecoli); enhances milk production and weight gain

     cellulase (ecoli) hydrolyzes cellulose making the whole plant good for feed

    Plants  easier to engineer – can be made from a single cell

    - helps with herbicide  resists pathogens and insects

    - enhanced food value 

15. Describe how plant genes can be manipulated using the Ti plasmid carried by Agrobacterium as a vector.

  • Ti plasmid = tumor-inducing

  • Agrobacterium is normally pathogenic

  • turned the plasmid into a useful vector by eliminating its disease-causing ability without interfering with its potential to move genetic materials into infected plants 

16. Explain how foreign DNA may be transferred into monocotyledonous plants.

  • Electroporation  high voltage jolts of electricity to open temporary pores in the cell membrane; foreign DNA can enter the cells through these pores

  • DNA particle gun  the DNA gun shoots tiny DNA-coated metal pellets through the cell walls into the cytoplasm, where foreign DNA becomes integrated into the host cell DNA 

17. Describe how recombinant DNA studies and the biotechnology industry are regulated with regards to safety and policy matters.

  • The processes could create hazardous new pathogens

  • Regulation begins with the self-monitoring approach

    - moves on to governments and regulatory agencies which promote potential industrial, medical, and agricultural benefits but ensure that the new products are safe 

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