Essentials of Genetics (9th Edition) - Standalone book
9th Edition
ISBN: 9780134047799
Author: William S. Klug, Michael R. Cummings, Charlotte A. Spencer, Michael A. Palladino
Publisher: PEARSON
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Chapter 17, Problem 15PDQ
Summary Introduction
To review:
In cDNA library formation, shorter cDNA clones are derived as compared to mRNA molecules.
Introduction:
cDNA Libraries are a collection of cDNA of one organism cloned into host cells. They are formed from mRNA isolated from cells. They only contain coding sequences and do not contain introns or other non-coding regions. Due to this reasons, cDNA libraries are preferred over genomic libraries. cDNA libraries are created by inserting vectors into cDNA and cloning to produce more copies.
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One procedure of obtaining cDNA from mRNA is by using oligo(dT) primers. What are oligo(dT)s? Why does using them make sense based on the processing (or modification) of precursor mRNA to get mature mRNA?
A protein has the following amino acid sequence:
Met-Tyr-Asn-Val-Arg-Val-Tyr-Lys-Ala-Lys-Trp-Leu-Ile-His-Thr-Pro
You wish to make a set of probes to screen a cDNA library for the sequence that encodes this protein. Your probes should be at least 18 nucleotides in length.
Q. How many different probes must be synthesized to be certain that you will find the cDNA sequence that specifies the protein?
The Reverse Transcriptase and Integrase enzymes produced by Retrotransposons sometimes mistakenly act on cellular mRNAs and generate a DNA intermediate that becomes integrated into the genome (which can lead to formation of new pseudogenes). Would you predict that a newly transposed element produced from an mRNA will generally be transcribed after its integration? Explain your answer.
Chapter 17 Solutions
Essentials of Genetics (9th Edition) - Standalone book
Ch. 17 -
CASE STUDY |Should we worry about recombinant DNA...Ch. 17 - Prob. 2CSCh. 17 - Prob. 3CSCh. 17 -
HOW DO WE KNOW?
1. In this chapter we focused on...Ch. 17 - Prob. 2PDQCh. 17 - What roles do restriction enzymes, vectors, and...Ch. 17 - Prob. 4PDQCh. 17 - Prob. 5PDQCh. 17 - Prob. 6PDQCh. 17 - Prob. 7PDQ
Ch. 17 - List the advantages and disadvantages of using...Ch. 17 - What are the advantages of using a restriction...Ch. 17 - The introduction of genes into plants is a common...Ch. 17 - Prob. 11PDQCh. 17 - Prob. 12PDQCh. 17 - Prob. 13PDQCh. 17 - What advantages do cDNA libraries provide over...Ch. 17 - Prob. 15PDQCh. 17 -
16. List the steps involved in screening a...Ch. 17 -
17. In a typical PCR reaction, describe what is...Ch. 17 -
18. We usually think of enzymes as being most...Ch. 17 - How are dideoxynucleotides (ddNTPs) structurally...Ch. 17 - Prob. 20PDQCh. 17 - One complication of making a transgenic animal is...Ch. 17 -
22. When disrupting a mouse gene by knockout, why...Ch. 17 - Prob. 23PDQ
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- What is the advantage of using a cDNA library?arrow_forwardAfter Drosophila DNA has been treated with a restriction enzyme, the fragments are inserted into plasmids and selected as clones in E. coli. With the use of this “shotgun” technique, every DNA sequence of Drosophila in a library can be recovered.a. How would you identify a clone that contains DNA encoding the protein actin, whose amino acid sequence is known?b. How would you identify a clone encoding a specific tRNA?arrow_forwardYou are studying a human gene, and try to express the protein in E. coli bacterial cells. To do this, you use lab cloning techniques to create an expression vector - a circular piece of DNA (plasmid) which can replicate within E. coli, and which contains an appropriate E. coli promoter sequence before the human protein sequence. Note that the sequence used is the final mRNA protein coding sequence, with all introns removed. You can detect that some of the protein is produced, but it's a very small amount. Luckily you included a positive control, which uses the same expression vector, and find that the control E. coli protein is expressed to high levels under the same conditions. A colleague suggests that you transform your same expression vector into a 'humanized' strain of E. coli that is optimized for the expression of human recombinant proteins (this is a real thing!). You do this and see protein expression – woohoo! If the genetic code is universal, why might a human gene be poorly…arrow_forward
- You isolate a mouse Tau-gene-containing DNA fragment from the chicken and hybridize it to the freshly-made and isolated hnRNA (primary transcript) from the nucleus of the mouse cells transcribed from the Tau gene (immediately after it was produced), allowing no time for processing of the hnRNA. Describe what you see when you look at the DNA/RNA hybrid molecule under the electron microscope.arrow_forwardWhat is the difference between a genomic library and a cDNA library?arrow_forwardYou want to clone the Human Gene A into a plasmid for producing the Protein A in bacteria. GeneA encodes an mRNA of 100 nucleotides long, but the entire gene spans more than 4000 nucleotides. There are three exons and two introns. If we were to clone this gene directly from the nuclear DNA, bacteria would not be able to express the insulin protein. Explain why this is true.arrow_forward
- Tryptophan synthase is one of the enzymes synthesized from the trp mRNA. In wild-type E. coli, the trp mRNA has a short half-life, but the tryptophan synthase half-life is much longer. To investigate how changes to the stability of the enzyme or its mRNA affect enzyme activity, two strains of E. coli were engineered. Strain A stabilizes the trp mRNA and strain B rapidly degrades tryptophan synthase. The wild-type, A, and B strains were grown in a medium with glucose as the sole carbon source. After several generations, tryptophan was added to all three cultures and tryptophan synthase activity was measured periodically. Note: Evaluate each condition as a simple model, where changes in the stability of trp mRNA or tryptophan synthase do not elicit secondary effects in the cells. Select the statements that describe the expected change in tryptophan synthase activity after the addition of tryptophan. In strain B, since both the trp mRNA and tryptophan synthase are rapidly degraded, the…arrow_forwardCould quantitative PCR, which uses a DNA-binding dye, to show how many copies of the target DNA sequence could be used to quantify the amount of mRNA in a cell? Would you expect that a metabolically active tissue such as the liver would show more cDNA copies in such a method, compared to less metabolically active tissues such as skin cells? One reason that the types and amounts of mRNAs are quantified in different tissue types is to compare which genes are activated and which are inactive. It used to be thought that any gene that was transcribed was automatically translated. The discovery of RNA-degrading systems shows that the real situation in cells is more complemented. Do you believe that a larger amount of mRNA of a given type, say for alpha hemoglobin in immature red blood cells is a reliable indicator that more alpha hemoglobin protein will be made in those cells?arrow_forwardA molecular geneticist hopes to find a gene in human liver cells that codes for an important blood-clotting protein. He knows that the nucleotife sequence of a small part of the gene is GTGGACTGACA. Briefly explain how to obtain the desired gene.arrow_forward
- A rice MADS box transcription factor gene contains a number of sequences that are removed in the processing of the mRNA transcript. Bacterial cells cannot excise these sequences from mRNA transcripts, yet via recombinant DNA technology, this gene can be cloned into a bacterial cell and produce the MADS box factor protein. Explain how this is possible.arrow_forwardSometimes, cDNAs turn out to be “chimeras”; that is, fusions of DNA copies of two different mRNAs accidentally inserted adjacently to each other in the same clone.You suspect that a cDNA clone from the nematodeCaenorhabditis elegans is such a chimera because the sequence of the cDNA insert predicts a protein with twostructural domains not normally observed in the sameprotein. How would you use the availability of the entiregenomic sequence to assess if this cDNA clone is a monster or not?arrow_forwardA molecular geneticist hopes to find a Gene in human liver cell that codes for an important blood-clotting protein,he knows that the nucleotide sequence of a small part of the Gene is GTGGACTGACA.briefly explain how to obtain genearrow_forward
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