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Biotechnology

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Biotechnology

So how does a scientist work with DNA?

It always starts with the sequence. Once the sequence is known, so much more can be done. Specific regions can be isolated, cloned, amplified, and then used to help us.

Biotechnology Methods

Biotechnology is the use of technology to change the genetic makeup of living things for human purposes. Generally, the purpose of biotechnology is to create organisms that are useful to humans or to cure genetic disorders. For example, biotechnology may be used to create crops that resist insect pests or yield more food, or to create new treatments for human diseases.

Biotechnology: The Invisible Revolution can be seen at http://www.youtube.com/watch?v=OcG9q9cPqm4 .

What does biotechnology have to do with me? is discussed in the following video: http://www.youtube.com/watch?v=rrT5BT_7HdI (10:01).

Biotechnology uses a variety of techniques to achieve its aims. Two commonly used techniques are gene cloning and the polymerase chain reaction.

Gene Cloning

Gene cloning is the process of isolating and making copies of a gene. This is useful for many purposes. For example, gene cloning might be used to isolate and make copies of a normal gene for gene therapy. Gene cloning involves four steps: isolation, ligation, transformation, and selection. You can watch an interactive animation about gene cloning at this link: http://www.teachersdomain.org/asset/biot09_int_geneclone/ .

  1. In isolation, an enzyme (called a restriction enzyme) is used to break DNA at a specific base sequence. This is done to isolate a gene.
  2. During ligation , the enzyme DNA ligase combines the isolated gene with plasmid DNA from bacteria. (A plasmid is circular DNA that is not part of a chromosome and can replicate independently.) Ligation is illustrated in Figure below . The DNA that results is called recombinant DNA .
  3. In transformation , the recombinant DNA is inserted into a living cell, usually a bacterial cell. Changing an organism in this way is also called genetic engineering .
  4. Selection involves growing transformed bacteria to make sure they have the recombinant DNA. This is a necessary step because transformation is not always successful. Only bacteria that contain the recombinant DNA are selected for further use.

Recombinant DNA

Ligation. DNA ligase joins together an isolated gene and plasmid DNA. This produces recombinant DNA.

Recombinant DNA technology is discussed in the following videos and animations: http://www.youtube.com/watch?v=x2jUMG2E-ic (4.36), http://www.youtube.com/watch?v=Jy15BWVxTC0 (0.50), http://www.youtube.com/watch?v=sjwNtQYLKeU (7.20), http://www.youtube.com/watch?v=Fi63VjfhsfI (3:59).

Polymerase Chain Reaction

The polymerase chain reaction (PCR) makes many copies of a gene or other DNA segment. This might be done in order to make large quantities of a gene for genetic testing. PCR involves three steps: denaturing , annealing , and extension . The three steps are illustrated in Figure below . They are repeated many times in a cycle to make large quantities of the gene. You can watch animations of PCR at these links:

  1. Denaturing involves heating DNA to break the bonds holding together the two DNA strands. This yields two single strands of DNA.
  2. Annealing involves cooling the single strands of DNA and mixing them with short DNA segments called primers . Primers have base sequences that are complementary to segments of the single DNA strands. As a result, bonds form between the DNA strands and primers.
  3. Extension occurs when an enzyme ( Taq polymerase or Taq DNA polymerase) adds nucleotides to the primers. This produces new DNA molecules, each incorporating one of the original DNA strands.

Polymerase chain reaction process

The Polymerase Chain Reaction. The polymerase chain reaction involves three steps. High temperatures are needed for the process to work. The enzyme Taq polymerase is used in step 3 because it can withstand high temperatures.

Summary

  • Biotechnology is the use of technology to change the genetic makeup of living things for human purposes.
  • Gene cloning is the process of isolating and making copies of a DNA segment such as a gene.
  • The polymerase chain reaction makes many copies of a gene or other DNA segment.

Practice

Use this resource and the videos associated with this resource to answer the questions that follow.

  1. Who developed PCR?
  2. What does PCR allow?
  3. Describe the 3 steps involved in PCR.
  4. Approximately how many copies of a specific segment of DNA can be made by PCR?

Review

  1. Define biotechnology.
  2. What is recombinant DNA?
  3. Identify the steps of gene cloning.
  4. What is the purpose of the polymerase chain reaction?
  5. Describe the three steps of PCR.

Vocabulary

annealing

annealing

Second step of the PCR process; involves binding of primers to denatured DNA strands.
biotechnology

biotechnology

Use of technology to change the genetic makeup of living things in order to produce useful products.
denaturing

denaturing

First step of the PCR process; involves heating DNA to break the bonds holding together the two DNA strands.
DNA ligase

DNA ligase

Enzyme that ligates (combines) two pieces of DNA together.
extension

extension

Third step of the PCR process; involves Taq polymerase adding nucleotides to the primers, producing new DNA molecules.
gene cloning

gene cloning

Process of isolating and making copies of a gene.
genetic engineering

genetic engineering

Using biotechnology to change the genetic makeup of an organism.
ligation

ligation

Process of combining two pieces of DNA together.
plasmid

plasmid

Small, circular piece of DNA in a prokaryotic cell.
polymerase chain reaction (PCR)

polymerase chain reaction (PCR)

Biotechnology process that makes many copies of a gene or other DNA segment.
primer

primer

Short segment of DNA; used to initiate extension by DNA polymerase during the PCR process.
recombinant DNA

recombinant DNA

DNA that results when DNA from two organisms is combined
Taq polymerase

Taq polymerase

An enzyme that adds nucleotides to the primers; also known as Taq DNA polymerase.
transformation

transformation

Changing an organism by inserting novel DNA into the organism; inserting recombinant DNA into a living cell, usually a bacterial cell.

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