Recombinant DNA Technology Overview
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Questions and Answers

What is the primary function of a vector in molecular biology?

  • To transport nutrients across cell membranes
  • To deactivate harmful genes in bacteria
  • To serve as a vehicle for the replication of desired DNA sequences (correct)
  • To produce proteins for cellular structures

    • Which feature is essential for a synthetic plasmid to initiate replication?

  • Multiple cloning site
  • Marker gene
  • Resistance gene
  • Origin of replication (ori) (correct)

    • What is the role of marker genes in synthetic plasmids?

  • To enhance the growth rate of bacteria
  • To provide a means of selecting transformed cells (correct)
  • To repair damaged DNA sequences
  • To identify the location of the plasmid within the cell

    • What distinguishes shuttle vectors from standard plasmids?

    <p>They can exist in multiple species and facilitate gene transfer (C)</p> Signup and view all the answers

    Why are resistance genes important in synthetic plasmids?

    <p>They help bacteria survive in selective environments, like those with antibiotics (D)</p> Signup and view all the answers

    What is the primary purpose of recombinant DNA technology?

    <p>To introduce a gene from one organism into a host cell (B)</p> Signup and view all the answers

    What enzyme class is responsible for cleaving DNA into fragments?

    <p>Restriction endonucleases (C)</p> Signup and view all the answers

    What must happen to the host cells that successfully uptake a DNA construct?

    <p>They must be identified and selected from other cells (C)</p> Signup and view all the answers

    Why are restriction enzymes significant in recombinant DNA technology?

    <p>They restrict the number of bacterial strains that can be infected (B)</p> Signup and view all the answers

    What is a cloning vector primarily used for in recombinant DNA technology?

    <p>To replicate inserted DNA in host cells (D)</p> Signup and view all the answers

    How can the efficiency of DNA uptake in host cells be characterized?

    <p>It is usually low, necessitating selective processes (A)</p> Signup and view all the answers

    What is formed when a DNA fragment is ligated to a cloning vector?

    <p>Recombinant DNA molecule (C)</p> Signup and view all the answers

    What is meant by 'recognition sequences' in the context of restriction endonucleases?

    <p>They are specific sequences where restriction enzymes cut the DNA (A)</p> Signup and view all the answers

    How do bacteria prevent their DNA from being degraded by endonucleases?

    <p>By modifying recognition sequences with methyl groups (D)</p> Signup and view all the answers

    What characterizes the recognition sequences of restriction endonucleases?

    <p>They read the same on both DNA strands in the 5' to 3' direction. (B)</p> Signup and view all the answers

    Which type of restriction enzyme is most commonly used in laboratory experiments?

    <p>Type II restriction enzymes (A)</p> Signup and view all the answers

    What is the main outcome of using DNA ligase in recombinant DNA technology?

    <p>It reattaches DNA sequences to create new DNA molecules. (C)</p> Signup and view all the answers

    What are the two types of ends produced by restriction enzymes when cutting DNA?

    <p>Sticky ends and blunt ends (C)</p> Signup and view all the answers

    What is a plasmid?

    <p>A small, circular piece of double-stranded DNA found in bacteria (A)</p> Signup and view all the answers

    Which of the following is a benefit provided by genes carried in plasmids?

    <p>Genetic advantages like antibiotic resistance (D)</p> Signup and view all the answers

    How do sticky ends of DNA fragments facilitate recombination?

    <p>By joining through hydrogen bonding and base pairing (B)</p> Signup and view all the answers

    Signup and view all the answers

    Study Notes

    Recombinant DNA Technology

    • Recombinant DNA technology, also known as gene cloning, genetic engineering, or molecular cloning, is a collection of protocols.
    • The goal of these manipulations is to introduce a gene from one organism into a host cell, where it replicates and is studied.
    • DNA is extracted from a source organism and treated enzymatically.
    • Fragments of DNA are joined to a cloning vector to form a recombinant DNA molecule (construct).
    • DNA constructs are transferred into a host cell.
    • Typically, a single DNA construct enters each host cell, though efficiency of uptake is low.
    • Host cells with the desired DNA construct need to be identified and separated from those without it.
    • The desired insert DNA of a construct can be characterized after isolation.
    • In some cases, the insert DNA is expressed in the host cell, and the encoded protein can be harvested.

    Restriction Endonucleases

    • These are DNA cutting enzymes found in bacteria.
    • Restriction enzymes limit the types of bacteriophages able to infect bacteria.
    • Each restriction enzyme recognizes a short, specific sequence of nucleotide bases.
    • These recognition sequences are called recognition sequences and are randomly spaced throughout the DNA.
    • Bacteria protect their own DNA from degradation by modifying their recognition sequences using methylases; these add methyl groups to adenine or cytosine bases within the recognition sequence.
    • Recognition sequences are often palindromic, meaning the sequence reads the same on both strands in 5´to 3´ direction.
    • Type II restriction enzymes are commonly used in laboratories as they cut DNA within their recognition sequence.

    Plasmids and Vectors

    • Plasmids are small, circular, double-stranded DNA molecules found in bacteria.
    • Plasmids exist independently from chromosomal DNA and replicate autonomously.
    • Plasmids provide bacteria with genetic advantages, such as antibiotic resistance.
    • Plasmids can range in size from a few thousand to hundreds of thousands of base pairs in length.
    • Synthetic plasmids are also called vectors in molecular biology.
    • Vectors are engineered plasmids, which serve as vehicles enabling replication of desired DNA sequences.
    • Vectors contain several characteristics that make them useful for molecular biologists including an origin of replication, a multiple cloning site, and selectable markers to enable identification of cells carrying the desired gene.

    Shuttle Vectors

    • Some plasmids can exist in multiple species.
    • These plasmids are called shuttle vectors.
    • Shuttle vectors can be used to move cloned DNA sequences among various organisms.
    • Shuttle vectors are useful for genetically modifying multicellular organisms, such as inserting herbicide resistance genes in plants.

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    Recombinant DNA Technology Lecture Notes PDF

    Description

    This quiz covers the fundamentals of recombinant DNA technology, including the processes of gene cloning and genetic engineering. Test your understanding of how DNA is manipulated and transferred into host cells, and the role of restriction endonucleases in these techniques.

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