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Biology: Restriction Enzymes and Nucleases
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Biology: Restriction Enzymes and Nucleases

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Questions and Answers

What is the primary goal of maintaining a sterile ambience in chemical engineering processes?

  • To preserve genetic information
  • To enable the growth of only the desired microbe or eukaryotic cell (correct)
  • To prevent contamination of microorganisms
  • To manufacture biotechnological products like antibiotics and vaccines

    • What is the major advantage of sexual reproduction over asexual reproduction?

  • It leads to inclusion and multiplication of undesirable genes
  • It preserves genetic information
  • It allows for variations and unique combinations of genetic setup (correct)
  • It is a faster process

    • What is the limitation of traditional hybridisation procedures?

  • They preserve genetic information
  • They lead to inclusion and multiplication of undesirable genes (correct)
  • They introduce only desirable genes
  • They are used in genetic engineering

    • What is the purpose of genetic engineering techniques?

    <p>To isolate and introduce only one or a set of desirable genes</p> Signup and view all the answers

    What happens to a piece of DNA transferred into an alien organism?

    <p>It gets integrated into the genome of the recipient and multiplies</p> Signup and view all the answers

    What is responsible for initiating replication in a chromosome?

    <p>The origin of replication</p> Signup and view all the answers

    What is the significance of gene cloning in genetic engineering?

    <p>It allows for the isolation and introduction of desirable genes</p> Signup and view all the answers

    What is the ultimate goal of genetic engineering?

    <p>To introduce desirable genes into an organism</p> Signup and view all the answers

    What is the purpose of having a single recognition site for restriction enzymes in a cloning vector?

    <p>To simplify the gene cloning process</p> Signup and view all the answers

    Which antibiotic resistance gene is commonly used as a selectable marker in E. coli?

    <p>All of the above</p> Signup and view all the answers

    What is the consequence of inserting foreign DNA at the BamH I site of the tetracycline resistance gene in the vector pBR322?

    <p>The recombinant plasmids will lose tetracycline resistance</p> Signup and view all the answers

    What is the function of the 'ori' region in the pBR322 vector?

    <p>It is involved in the replication of the plasmid</p> Signup and view all the answers

    Why are normal E. coli cells not resistant to antibiotics?

    <p>They do not carry resistance genes</p> Signup and view all the answers

    What is the purpose of plating transformants on ampicillin-containing medium?

    <p>To select against non-recombinant plasmids</p> Signup and view all the answers

    What is the name of the restriction enzyme site where foreign DNA is commonly ligated in the vector pBR322?

    <p>BamH I</p> Signup and view all the answers

    What is the advantage of using a cloning vector with antibiotic resistance genes?

    <p>It allows for the selection of recombinant plasmids</p> Signup and view all the answers

    What does the letter 'R' in EcoRI represent?

    <p>Name of the strain of bacteria</p> Signup and view all the answers

    What is the function of exonucleases?

    <p>They remove nucleotides from the ends of the DNA</p> Signup and view all the answers

    What is the characteristic of a palindrome in DNA?

    <p>It is a sequence of base pairs that reads same on the two strands</p> Signup and view all the answers

    What is the purpose of restriction endonuclease in DNA?

    <p>To make cuts at specific positions within the DNA</p> Signup and view all the answers

    What is the general name of the class of enzymes that restriction endonuclease belongs to?

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

    What is the recognition sequence of a restriction endonuclease?

    <p>A specific palindromic nucleotide sequence</p> Signup and view all the answers

    What is the direction of reading in a palindrome in DNA?

    <p>5' to 3' direction</p> Signup and view all the answers

    What is the role of restriction endonuclease in forming recombinant DNA?

    <p>It makes cuts at specific positions within the DNA</p> Signup and view all the answers

    What happens to the antibiotic resistance gene when alien DNA is inserted?

    <p>It gets inactivated</p> Signup and view all the answers

    What is the purpose of using two antibiotics in the selection process?

    <p>To differentiate between recombinants and non-recombinants</p> Signup and view all the answers

    What is the result of insertional inactivation of β-galactosidase?

    <p>No production of colour</p> Signup and view all the answers

    What is the purpose of using a chromogenic substrate?

    <p>To differentiate between recombinants and non-recombinants</p> Signup and view all the answers

    What type of colonies are produced when the plasmid has an insert?

    <p>Colourless colonies</p> Signup and view all the answers

    What is the result of inserting a recombinant DNA within the coding sequence of β-galactosidase?

    <p>Inactivation of β-galactosidase</p> Signup and view all the answers

    What is the advantage of using alternative selectable markers?

    <p>It is simpler and more efficient</p> Signup and view all the answers

    What happens when a plasmid does not have an insert?

    <p>It produces blue colour</p> Signup and view all the answers

    What is necessary to increase the efficiency of DNA entering a bacterium?

    <p>A specific concentration of a divalent cation</p> Signup and view all the answers

    What is the purpose of heat shock in introducing recombinant DNA into bacteria?

    <p>To allow the bacteria to take up the recombinant DNA</p> Signup and view all the answers

    What method is used to introduce recombinant DNA into plant cells?

    <p>Biolistics or gene gun</p> Signup and view all the answers

    What is the first step in recombinant DNA technology?

    <p>Isolation of DNA</p> Signup and view all the answers

    What is the purpose of ligation in recombinant DNA technology?

    <p>To join the DNA fragment into a vector</p> Signup and view all the answers

    What is the genetic material of all organisms?

    <p>Deoxyribonucleic acid or DNA</p> Signup and view all the answers

    Why is it necessary to have DNA in pure form, free from other macro-molecules?

    <p>To cut the DNA with restriction enzymes</p> Signup and view all the answers

    What is the final step in recombinant DNA technology?

    <p>Extraction of the desired product</p> Signup and view all the answers

    Study Notes

    Restriction Enzymes

    • EcoRI is a restriction enzyme with 'R' derived from the name of the strain of bacteria it was isolated from
    • Restriction enzymes are a type of nucleases that make cuts at specific positions within the DNA
    • Endonucleases, like restriction enzymes, cut DNA at specific positions, whereas exonucleases remove nucleotides from the ends of the DNA

    Palindromic Sequences

    • Palindromes are groups of letters that form the same words when read both forward and backward
    • In DNA, palindromic sequences are sequences of base pairs that read the same on the two strands when orientation of reading is kept the same

    Genetic Engineering

    • Genetic engineering involves creation of recombinant DNA, use of gene cloning, and gene transfer
    • This technique allows for the isolation and introduction of only one or a set of desirable genes without introducing undesirable genes into the target organism
    • Traditional hybridization procedures in plant and animal breeding often lead to inclusion and multiplication of undesirable genes along with the desired genes

    Gene Cloning

    • Cloning sites in vectors need to have very few, preferably single, recognition sites for commonly used restriction enzymes
    • Presence of more than one recognition site within the vector will generate several fragments, which will complicate the gene cloning process

    Selectable Markers

    • Genes encoding resistance to antibiotics such as ampicillin, chloramphenicol, tetracycline, or kanamycin are considered useful selectable markers for E. coli
    • Normal E. coli cells do not carry resistance against any of these antibiotics
    • Alternative selectable markers have been developed which differentiate recombinants from non-recombinants on the basis of their ability to produce color in the presence of a chromogenic substrate

    Recombinant DNA Technology

    • Recombinant DNA technology involves several steps in a specific sequence such as isolation of DNA, fragmentation of DNA by restriction endonucleases, isolation of a desired DNA fragment, ligation of the DNA fragment into a vector, transferring the recombinant DNA into the host, culturing the host cells in a medium at large scale, and extraction of the desired product

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    Description

    This quiz is about restriction enzymes, specifically EcoRI, and their relationship to nucleases. It covers the origin of the name 'R' in EcoRI and the classification of these enzymes.

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