Recombinant DNA Technology Tools
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Questions and Answers

What specific sequence does EcoRI recognize for cutting DNA?

  • GAATTC (correct)
  • AGCT
  • GATC
  • CTTAAG

What is formed at the ends of DNA fragments after being cut by restriction enzymes like EcoRI?

  • Circular ends
  • Sticky ends (correct)
  • Blunt ends
  • Double-stranded ends

What role does DNA ligase play in the process of recombinant DNA formation?

  • It reads the DNA sequence.
  • It cuts the DNA strands.
  • It facilitates base pairing.
  • It joins DNA fragments together. (correct)

Which of the following best describes a DNA palindrome?

<p>A sequence that reads the same in both directions. (B)</p> Signup and view all the answers

Which of the following statements about restriction enzymes is incorrect?

<p>They join DNA fragments using hydrogen bonds. (A)</p> Signup and view all the answers

What term describes the DNA molecules composed of DNA from different sources?

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

In creating recombinant DNA, why is it important for the DNA fragments to have the same sticky ends?

<p>To ensure they bind with high affinity. (C)</p> Signup and view all the answers

How do restriction enzymes like EcoRI contribute to genetic engineering?

<p>They facilitate the insertion of foreign DNA into host organisms. (A)</p> Signup and view all the answers

What is a primary function of restriction endonucleases in recombinant DNA technology?

<p>To cut DNA at specific points (A)</p> Signup and view all the answers

Which characteristic defines the recognition sequence for restriction enzymes?

<p>It is always palindromic. (B)</p> Signup and view all the answers

What distinguishes endonucleases from exonucleases?

<p>They cut DNA at specific internal locations. (C)</p> Signup and view all the answers

Which of the following is an example of a restriction enzyme?

<p>EcoRI (D)</p> Signup and view all the answers

What type of research led to the isolation of restriction enzymes in 1963?

<p>Bacteriophage studies (C)</p> Signup and view all the answers

How are restriction enzymes typically named?

<p>Based on their bacterial origin and isolation order (B)</p> Signup and view all the answers

What role do restriction enzymes play in the process of genetic engineering?

<p>They recognize and cut specific DNA sequences. (A)</p> Signup and view all the answers

Which of the following statements is true about the number of restriction enzymes known today?

<p>There are approximately 900 restriction enzymes known. (B)</p> Signup and view all the answers

What is the role of restriction endonucleases in recombinant DNA technology?

<p>They cut DNA at specific points. (B)</p> Signup and view all the answers

Why is agarose gel used in gel electrophoresis?

<p>It acts as a natural polymer that helps separate DNA fragments. (D)</p> Signup and view all the answers

What happens to DNA fragments during gel electrophoresis?

<p>They are separated by size when an electric field is applied. (A)</p> Signup and view all the answers

How can separated DNA fragments be visualized after electrophoresis?

<p>By staining with ethidium bromide and exposing to UV radiation. (B)</p> Signup and view all the answers

What is the implication of cutting both the vector and foreign DNA with the same restriction enzyme?

<p>It allows the joining of DNA fragments to create a recombinant vector. (A)</p> Signup and view all the answers

The movement of DNA fragments in agarose gel electrophoresis is influenced most significantly by which factor?

<p>The size of the DNA fragments. (B)</p> Signup and view all the answers

During transformation in recombinant DNA technology, what is the end result?

<p>Foreign DNA is integrated into bacterial cells. (B)</p> Signup and view all the answers

What characteristic of DNA fragments allows them to be separated using gel electrophoresis?

<p>Their size and negative charge. (D)</p> Signup and view all the answers

Flashcards

Restriction Enzymes

Enzymes that cut DNA molecules at specific sequences called recognition sequences.

Recognition Sequence

A specific sequence of DNA nucleotides that a restriction enzyme recognizes and cuts.

Hind II

A specific restriction enzyme that recognizes a 6 base pair sequence.

EcoRI

A restriction enzyme isolated from Escherichia coli strain RY13.

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Exonucleases

Enzymes that remove nucleotides from the ends of DNA.

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Endonucleases

Enzymes that cut DNA within the molecule at specific sequences.

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Palindromic Nucleotide Sequences

DNA sequences that read the same forwards and backwards on opposite strands.

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How do restriction enzymes work?

They 'inspect' DNA for their recognition sequences and then cut both strands of the DNA at specific points.

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Recombinant DNA

DNA created by combining genetic material from different sources.

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Sticky Ends

Complementary single-stranded DNA overhangs created by restriction enzymes.

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Ligase

An enzyme that joins DNA fragments together by forming phosphodiester bonds.

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Transformation

The process of introducing foreign DNA into a host cell.

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Gel Electrophoresis

A technique used to separate DNA fragments based on size.

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Ethidium Bromide

A fluorescent dye that binds to DNA and allows visualization under UV light.

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Agarose

A natural polymer extracted from seaweed, used as a matrix in gel electrophoresis.

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What are palindromes in DNA?

Palindromic DNA sequences read the same forwards and backwards on opposite strands, like the word 'MADAM'.

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How do restriction enzymes cut DNA?

They cut both DNA strands at specific points within a palindromic sequence, leaving single-stranded overhangs called 'sticky ends'.

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What are sticky ends?

Single-stranded overhangs created by restriction enzymes, which can base-pair with complementary sticky ends from other DNA fragments.

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What is recombinant DNA?

DNA created by combining genetic material from different sources; formed by joining DNA fragments with compatible sticky ends.

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What is the role of DNA ligase?

DNA ligase joins DNA fragments together at their sticky ends, forming a continuous DNA molecule.

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What is the purpose of restriction enzymes in genetic engineering?

Restriction enzymes cut DNA at specific sites, allowing us to insert foreign genes into vectors and create recombinant DNA molecules.

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How do sticky ends facilitate DNA joining?

Sticky ends, with their complementary sequences, allow DNA fragments cut by the same restriction enzyme to easily join together.

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What is the significance of the same restriction enzyme being used?

Using the same restriction enzyme ensures that the DNA fragments have compatible sticky ends, allowing them to combine.

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Study Notes

Recombinant DNA Technology Tools

  • Genetic engineering/recombinant DNA technology requires specific tools: restriction enzymes, polymerase enzymes, ligases, vectors, and host organisms
  • Restriction enzymes are crucial for cutting DNA at specific points
  • 1963: Enzymes responsible for restricting bacteriophage growth in Escherichia coli were isolated. One added methyl groups, the other cut DNA, called restriction endonuclease.
  • Hind II (restriction endonuclease) functions depend on specific DNA nucleotide sequence; It was isolated and characterized five years after.
  • Hind II cuts DNA at specific point recognizing a 6-base-pair sequence
  • Over 900 restriction enzymes are known now, isolated from >230 bacteria strains, each recognizing different sequences.
  • Enzyme naming convention: initials of genus followed by species name. e.g., EcoRI from Escherichia coli RY13

Restriction Enzymes - Detail

  • Enzymes recognize specific palindromic DNA sequences.
  • These sequences read the same on both strands when read 5' to 3' or 3' to 5' directions. Example: GAATTC/CTTAAG.
  • Enzymes cut both DNA strands at specific points within the palindrome sequence, creating "sticky ends"
  • Sticky ends are single-stranded portions.
  • Overhanging/ sticky ends easily bond with complementary sequences (hydrogen bonds) leading to recombination.
  • Restriction enzymes are used in genetic engineering to combine DNA fragments from different sources.

Restriction Enzyme Action

  • Exonucleases remove nucleotides from DNA ends
  • Endonucleases cut DNA at specific internal positions.
  • Restriction enzymes recognize a specific 6-base pair sequence within a DNA molecule and cut at a specific point to create sticky ends allowing the joining of fragments together
  • Different DNA fragments with compatible sticky ends can be joined from different species sources using DNA ligase.

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Biotechnology - Recombinant DNA Technology PDF

Description

This quiz explores the essential tools used in recombinant DNA technology, focusing on restriction enzymes and their functionalities. Learn about the history, naming conventions, and the importance of these enzymes in genetic engineering. Test your understanding of how these tools facilitate the manipulation of DNA.

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