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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

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?

The U.S. Department of Justice has established a database that catalogs \(P C R\) amplification products from short tandem repeats of the \(Y\) (Y-SIRs) chromosome in humans. The database contains polymorphisms of five U.S. ethnic groups (African Americans, European Americans, Hispanics, Native Americans, and Asian Americans) as well as worldwide population (a) Given that STRs are repeats of varying lengths, for example \((\mathrm{TCTG})_{9-17}\) or \((\mathrm{TAT})_{6-14}\) explain how PCR could reveal differences (polymorphisms) among individuals. How could the Department of Justice make use of those differences? (b) \(Y-S T R s\) from the nonrecombining region of the \(Y\) chromosome (NRY) have special relevance for forensic purposes. Why? (c) What would be the value of knowing the ethnic population differences for \(Y\) -STR polymorphisms? (d) For forensic applications, the probability of a "match" for a crime scene DNA sample and a suspect's DNA often culminates in a guilty or innocent verdict. How is a "match" determined, and what are the uses and limitations of such probabilities?

Describe the Cre-Lox system for generating conditional knockout mice. What type of genes can be studied by using an inducible promoter to activate the Cre gene?

The introduction of genes into plants is a common practice that has generated not only a host of genetically modified foodstuffs, but also significant worldwide controversy. Interestingly, a tumor-inducing plasmid is often used to produce genetically modified plants. Is the use of a tumor-inducing plasmid the source of such controversy?

If you performed a PCR experiment starting with only one copy of double- stranded DNA, approximately how many DNA molecules would be present in the reaction tube after 15 cycles of amplification?
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