Imagine you want to study one of the human crystallins, proteins present in the lens of the eye (see Figure 1.8). To obtain a sufficient amount of the protein of interest, you decide to clone the gene that codes for it. Assume you know the sequence of this gene. Explain how you would go about this.

Short Answer

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When the nucleotide sequence of a gene is known, this gene can be amplified by using a polymerase chain reaction that produces billions of copies of DNA in a few hours.

The primers can be taken that are complementary to the sequence of gene coding crystalline protein to attach the gene, and then many cycles of PCR can produce a large number of genes that will code for crystalline protein.

Step by step solution

01

Protein

Proteins are types of biomolecules formed after polymerization in amino acid molecules. All proteins variate in their sequence of amino acids that are coded by the DNA sequence of a gene. Their activity is added after the suitable post-translation modification in the cells.

02

Step 2:Crystallin

Crystallins are structural proteins in the lens of human eyes that are found outside of the lens. Two gene families are found that are alpha-crystallins and betagamma-crystallins. Alpha-crystallins work as chaperones that assist in proper protein interactions.

03

Step 3: Cloning of the crystalline gene

According to the given scenario, the sequence of the gene to be cloned is known. Thus, many copies of the gene are produced in the lab by using polymerase chain reaction, a technique used to amplify a gene and make billions of copies in a short duration.

A segment of the target gene sequence, Taq DNA polymerase, nucleotides, and primers that have complementary sequences of the target sequence are used in PCR reactions. Primers help to amplify the desired sequence by repeated cycles of denaturation, annealing, and extension in PCR.

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Most popular questions from this chapter

You are cloning an aardvark gene, using a bacterial plasmid as a vector. The green diagram shows the plasmid, which contains the restriction site for the enzyme used in Figure 20.5. Above the plasmid is a segment of linear aardvark DNA that was synthesized using PCR. Diagram your cloning procedure, and show what would happen to these two molecules during each step. Use one color for the aardvark DNA and its bases and another color for those of the plasmid. Label each step and all 5’ and 3’ ends.

You hope to study a gene that codes for a neurotransmitter protein produced in human brain cells. You know the amino acid sequence of the protein. Explain how you might (a) identify what genes are expressed in a specific type of brain cell, (b) identify (and isolate) the neurotransmitter gene, (c) produce multiple copies of the gene for study, and(d) produce large quantities of the neurotransmitter for evaluation as a potential medication.

Which of the following tools of DNA technology is incorrectly paired with its use?

(A) electrophoresis—separation of DNA fragments

(B) DNA ligase—cutting DNA, creating sticky ends of restriction

fragments

(C) DNA polymerase—polymerase chain reaction to amplify

sections of DNA

(D) reverse transcriptase—production of cDNA from

mRNA

What is the advantage of using stem cells for gene therapy or gene editing?

If the template strand has two or more identical nucleotides in a row, their complementary nucleotides will be added one after the other in the same flow step. How are two or more of the same nucleotide (in a row) detected in the flowgram? (See sample on the right.) Write out the sequence of the first 25 nucleotides in the flow-gram above, starting from the left (Ignore the very short lines.)

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