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Chapter 20: Problem 11

What is a cDNA library, and for what purpose can it be used?

Short Answer

Expert verified
Answer: A cDNA library is a collection of complementary DNA (cDNA) molecules derived from the messenger RNA (mRNA) molecules found in a specific tissue or organism. This library preserves the original gene expression pattern of the source material and can be used for various molecular biology and genomic research purposes. Applications of a cDNA library include gene expression studies, cloning genes, functional genomics, and sequencing.

Step by step solution

01

Introduction

A cDNA library is a collection of complementary DNA (cDNA) molecules derived from the messenger RNA (mRNA) molecules found in a specific tissue or organism. This library preserves the original gene expression pattern of the source material and can be used for various molecular biology and genomic research purposes.
02

cDNA synthesis

The first step in creating a cDNA library is synthesizing cDNA from the mRNA molecules. This process is called reverse transcription. An enzyme called reverse transcriptase specifically binds to the mRNA template, recognizes the poly-T tail, and synthesizes a single-stranded cDNA molecule. This single-stranded cDNA is further converted into a double-stranded cDNA using DNA polymerase.
03

cDNA Cloning

Once double-stranded cDNA is synthesized, it's cloned into a suitable vector (such as a plasmid) using restriction enzymes and DNA ligase. The vector containing the cDNA is then introduced into a host organism, typically a bacterial cell, which helps amplify and replicate the cDNA.
04

Library Screening

After cloning, the cDNA library can be screened for the desired cDNA. Various methods, such as nucleic acid hybridization and polymerase chain reaction (PCR), are used to identify the cDNA of interest from the library.
05

Applications of cDNA library

The cDNA library is used for numerous purposes: 1. Gene expression studies: By comparing cDNA libraries of different tissues or organisms, researchers can determine the patterns of gene expression. 2. Cloning genes: A cDNA library can serve as a source of specific genes for cloning and further analysis. 3. Functional genomics: cDNA libraries provide crucial information on gene function and regulation in the source tissue or organism. 4. Sequencing: cDNA sequences can be determined and compared to genomic sequences to identify exons and introns within the genes.

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Most popular questions from this chapter

What is the difference between a knockout animal and a transgenic animal?

In this chapter we focused on how specific DNA sequences can be copied, identified, characterized, and sequenced. At the same time, we found many opportunities to consider the methods and reasoning underlying these techniques. From the explanations given in the chapter, what answers would you propose to the following fundamental questions? (a) In a recombinant DNA cloning experiment, how can we determine whether DNA fragments of interest have been incorporated into plasmids and, once host cells are transformed, which cells contain recombinant DNA? (b) When using DNA libraries to clone genes, what combination of techniques are used to identify a particular gene of interest? (c) What steps make \(P C R\) a chain reaction that can produce millions of copies of a specific DNA molecule in a matter of hours without using host cells? (d) How has DNA sequencing technology evolved in response to the emerging needs of genome scientists?

The human insulin gene contains a number of sequences that are removed in the processing of the mRNA transcript. In spite of the fact that bacterial cells cannot excise these sequences from mRNA transcripts, explain how a gene like this can be cloned into a bacterial cell and produce insulin.

To create a cDNA library, cDNA can be inserted into vectors and cloned. In the analysis of \(\mathrm{cDNA}\) clones, it is often difficult to find clones that are full length-that is, many clones are shorter than the mature mRNA molecules from which they are derived. Why is this so?

One complication of making a transgenic animal is that the transgene may integrate at random into the coding region, or the regulatory region, of an endogenous gene. What might be the consequences of such random integrations? How might this complicate genetic analysis of the transgene?
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