The Basics of Recombinant DNA
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Questions and Answers

What is the main difference between microinjection and biolistic transformation?

  • The presence of bacteria in the process
  • The method of DNA delivery (correct)
  • The type of DNA being injected
  • The type of host cell used
  • What is the primary purpose of phage introduction in genetic engineering?

  • To use lambda or MI3 phages for transformation
  • To produce phage plaques
  • To identify recombinants from non-recombinants (correct)
  • To transfect cells with phages instead of bacteria
  • Who developed methods for splitting DNA molecules at selected sites and attaching segments to the DNA of a virus or plasmid?

  • Hamilton O. Smith
  • Stanley N. Cohen and Herbert W. Boyer
  • Paul Berg (correct)
  • Werner Arber
  • What is the fundamental goal of laboratory geneticists?

    <p>To isolate, characterize, and manipulate genes</p> Signup and view all the answers

    Which technology forms the basis of recombinant DNA technology?

    <p>Cloning and DNA sequencing</p> Signup and view all the answers

    What is the process of transfection in genetic engineering equivalent to?

    <p>Transformation using phages</p> Signup and view all the answers

    What is another term for Recombinant DNA as mentioned in the text?

    <p>Chimera</p> Signup and view all the answers

    Which process involves combining the DNA of two different organisms?

    <p>Transformation</p> Signup and view all the answers

    What is the purpose of a selectable marker in Recombinant DNA?

    <p>To identify recombinant molecules</p> Signup and view all the answers

    Which enzyme is used to cut a piece of DNA during the process of making Recombinant DNA?

    <p>Restriction Enzyme</p> Signup and view all the answers

    What happens to a host cell without a vector when exposed to a certain antibiotic?

    <p>It dies</p> Signup and view all the answers

    Which organism is given as an example of a possible host cell in the text?

    <p>E.Coli</p> Signup and view all the answers

    Study Notes

    Recombinant DNA Basics

    • Recombinant DNA is a general term for combining a piece of one DNA with another strand of DNA, also referred to as "chimera".
    • The most common recombinant process involves combining the DNA of two different organisms.

    Recombinant DNA Methods

    • There are three methods to make Recombinant DNA: Transformation, Phage Introduction, and Non-Bacterial Transformation.

    Transformation Method

    • Select a piece of DNA to be inserted into a vector.
    • Cut the DNA with a restriction enzyme.
    • Ligate the DNA insert into the vector with DNA Ligase.
    • The inserted DNA contains a selectable marker, often an antibiotic marker, to identify recombinant molecules.
    • The vector is inserted into a host cell, which is a process called transformation.
    • E.Coli is an example of a possible host cell.

    Non-Bacterial Transformation Method

    • In microinjection, the DNA is injected directly into the nucleus of the cell being transformed.
    • In Biolistic, the host cells are bombarded with high-velocity micro-projectiles, such as particles of gold or tungsten, coated with DNA.
    • Non-bacterial transformation does not use bacteria such as E.Coli as the host.

    Phage Introduction Method

    • In vitro packaging of a vector is used to produce phage plaques containing recombinant DNA.
    • Lambda or MI3 phages are used to produce phage plaques.
    • Recombinants can be identified by differences in the recombinants and non-recombinants using various selection methods.

    Historical Background

    • Werner Arber, a Swiss microbiologist, discovered restriction enzymes in 1968.
    • Hamilton O. Smith purified Type II restriction enzymes in 1969.
    • Paul Berg developed methods for splitting DNA molecules at selected sites and attaching segments of the molecule to the DNA of a virus or plasmid.
    • Stanley N. Cohen and Herbert W. Boyer were the first to insert recombined genes into bacterial cells in 1973.

    Background Information

    • Recombinant DNA technology is the joining of DNA molecules from two different species to produce new genetic combinations.
    • The goal of laboratory geneticists is to isolate, characterize, and manipulate genes.
    • Recombinant DNA technology is based on cloning and DNA sequencing.
    • Cloning is undertaken to obtain the clone of one particular gene or DNA sequence of interest.

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    Description

    Learn about the fundamentals of Recombinant DNA, including its process and applications. This quiz covers topics like combining DNA strands from different organisms and methods for creating Recombinant DNA.

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