Ch. 12 DNA Technology and Genomics

Taylor - Campbell Biology: Concepts & Connections 10th Edition

Taylor, Simon, Dickey, Hogan10th EditionCampbell Biology: Concepts & ConnectionsISBN: 9780136538783Not the one you use?Change textbook

Taylor, Simon, Dickey, Hogan 10th Edition

Ch. 12 DNA Technology and Genomics

Problem 9

Chapter 12, Problem 9

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.

Verified step by step guidance

Identify a sample source: The biochemist should start by obtaining a sample of human cells that are likely to express the blood-clotting protein, such as blood cells.

Extract DNA: From the obtained cells, extract the total DNA which contains the genetic information of the cells.

Use PCR to amplify the gene: Utilize the Polymerase Chain Reaction (PCR) technique to amplify the specific gene segment. Design primers that match the known nucleotide sequence (CTGGACTGACA) to target and amplify the gene of interest.

Clone and sequence the gene: Insert the amplified gene into a plasmid and introduce it into bacteria for cloning. Once cloned, sequence the DNA to confirm that the correct gene has been isolated and to determine the full sequence of the gene.

Verify the protein function: After confirming the gene sequence, express the protein in a suitable system and perform tests to verify that it functions as the blood-clotting protein.

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Key Concepts

Here are the essential concepts you must grasp in order to answer the question correctly.

Gene Isolation Techniques

Gene isolation involves methods to extract a specific gene from a genome. Techniques such as polymerase chain reaction (PCR) can amplify the target gene, making it easier to study. This process typically requires knowledge of the gene's nucleotide sequence, which allows for the design of specific primers that bind to the gene of interest.

Nucleotide Sequence and Primers

A nucleotide sequence is the order of nucleotides in a DNA strand, which encodes genetic information. In this case, the sequence CTGGACTGACA can be used to design primers—short DNA fragments that initiate DNA synthesis during PCR. These primers are crucial for selectively amplifying the desired gene from the complex mixture of DNA in human cells.

Cloning Vectors

Cloning vectors are DNA molecules used to transport foreign genetic material into a host cell for replication. After isolating the desired gene, it can be inserted into a vector, such as a plasmid, which can then be introduced into bacterial or eukaryotic cells. This allows for the production and study of the blood-clotting protein in a controlled environment.

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Cloning with Recombinant DNA

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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

Why does DNA profiling rely on comparing specific genetic markers rather than the entire genome?

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Textbook Question

Recombinant DNA techniques are used to custom-build bacteria for two main purposes: to obtain multiple copies of certain genes and to obtain useful proteins produced by certain genes. Give an example of each of these applications in medicine and agriculture.

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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

Explain how you might engineer E. coli to produce human growth hormone (HGH) using the following: E. coli containing a plasmid, DNA carrying the gene for HGH, DNA ligase, a restriction enzyme, equipment for manipulating and growing bacteria, a method for extracting and purifying the hormone, and an appropriate DNA probe. (Assume that the human HGH gene lacks introns.)

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Textbook Question

What is left for genetic researchers to do now that the Human Genome Project has determined nearly complete nucleotide sequences for all of the human chromosomes? Explain.

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