18. Biotechnology

Steps to DNA Cloning

18. Biotechnology

Steps to DNA Cloning

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Step 1) Create Recombinant DNA

1a) Use Restriction Enzymes

1b) Use Ligation Enzymes

2) Transform Recombinant DNA into Bacteria

Review & Application of DNA Cloning in Medicine

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01:16

BioInteractive: Genetic Engineering

Animation: Restriction Enzymes

Animation: Creating Recombinant DNA

Key Steps of Molecular Cloning

Mechanism of Recombination

[LECT C8 : RECOMBINANT DNA TECHNOLOGY] 8.2 Steps in Gene Cloning

Introduction to Molecular Cloning

DNA cloning and recombinant DNA | Biomolecules | MCAT | Khan Academy

Khan Academy

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Practice this topic

Multiple Choice

Human DNA cut with restriction enzyme A can be joined to:

Each restriction enzyme has a specific sequence of nucleotides where it cuts the DNA. These sequences of DNA are unique to each restriction enzyme and are known as:

The single-stranded ends of DNA molecules can be joined together by:

The process of using DNA from one organism to alter the characteristics of another is called:

An organism which has foreign genes incorporated into its genomes is known as a:

What is the most logical sequence of steps for splicing foreign DNA into a plasmid and inserting the plasmid into a bacterium?
I. Transform bacteria with a recombinant DNA molecule.
II. Cut the plasmid DNA using restriction enzymes (endonucleases).
III. Extract plasmid DNA from bacterial cells.
IV. Hydrogen-bond the plasmid DNA to non-plasmid DNA fragments. V. Use ligase to seal plasmid DNA to non-plasmid DNA.

What two enzymes are needed to produce recombinant DNA?

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Multiple Choice

Which arrangement of the following four enzymes represents the order in which they would be used in a typical gene-cloning experiment resulting in the insertion of a cDNA into a bacterial plasmid? Begin with the gene's mRNA transcript.

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Multiple Choice

What is the enzymatic function of restriction enzymes?

What do restriction endonucleases do?

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Textbook Question

What do restriction endonucleases do?

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Textbook Question

What is a plasmid? a. an organelle found in many bacteria and certain eukaryotes b. a circular DNA molecule that replicates independently of the main chromosome(s) c. a type of virus that has a DNA genome and infects certain types of human cells, including lung and respiratory tract tissue d. a type of virus that has an RNA genome, codes for reverse transcriptase, and inserts a cDNA copy of its genome into cells

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Textbook Question

After finding a gene that causes a disease, researchers often introduce the defective allele into mice to create an animal model of the disease. Why are these models valuable? a. They allow the testing of potential drug therapies without endangering human patients. b. They allow the sequencing of the mutant allele. c. They allow the production of large quantities of the defective gene product, usually a protein. d. They allow the study of how the gene was transmitted from parents to offspring.

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Textbook Question

DNA technology has many medical applications. Which of the following is not done routinely at present? a. production of hormones for treating diabetes and dwarfism b. analysis of gene expression for more informed cancer treatments c. gene editing by the CRISPR-Cas9 system in viable human embryos to correct genetic diseases d. prenatal identification of genetic disease alleles

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Textbook Question

When a typical restriction enzyme cuts a DNA molecule, the cuts are uneven, giving the DNA fragments single-stranded ends. These ends are useful in recombinant DNA work because a. they enable a cell to recognize fragments produced by the enzyme. b. they serve as starting points for DNA replication. c. the fragments will bond to other fragments with complementary ends. d. they enable researchers to use the fragments as molecular probes.

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Textbook Question

Expression of a cloned eukaryotic gene in a bacterial cell involves many challenges. The use of mRNA and reverse transcriptase is part of a strategy to solve the problem of a. post-transcriptional processing. b. post-translational processing. c. nucleic acid hybridization. d. restriction fragment ligation.

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Textbook Question

Which of the following sequences in double-stranded DNA is most likely to be recognized as a cutting site for a restriction enzyme? a. AAGG TTCC b. GGCC CCGG c. ACCA TGGT d. AAAA TTTT

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Textbook Question

A biochemist hopes to find a gene in human cells that codes for an important blood-clotting protein. She knows that the nucleotide sequence of a small part of the blood-clotting gene is CTGGACTGACA. Briefly outline a possible method she might use to isolate the desired gene.

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Textbook Question

A biologist isolated a gene from a human cell, inserted it into a plasmid, and inserted the plasmid into a bacterium. The bacterium made a new protein, but it was nothing like the protein normally produced in a human cell. Why? (Explain your answer.) a. The bacterium had undergone transformation. b. The gene did not have sticky ends. c. The human gene contained introns. d. The gene was not synthesized from scratch.

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Textbook Question

Potato blight causes potato plants to shrivel and rot. The disease is caused by the pathogen Phytophthora infestans, infamous for its role in Ireland's Great Potato Famine in the mid-1840s. The disease can devastate crops during wet weather, sometimes leading to total crop loss. Researchers aim to use recombinant DNA methods to transfer blight resistance genes from resistant varieties into susceptible varieties of potato. Explain how restriction endonucleases and DNA ligase could be used to insert a potato blight resistance gene into a plasmid.