EBK CONCEPTS OF GENETICS
12th Edition
ISBN: 9780134818979
Author: Killian
Publisher: YUZU
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Chapter 12, Problem 21ESP
Summary Introduction
To explain: The nucleosome-DNA interaction and gene activity based on the study related to in-vivo chemical modification of histones in yeast.
Introduction: Nucleosome is a basic unit for DNA packaging of a eukaryotic chromosome. It consists of a segment of DNA wrapped around an octamer of histone core proteins. It is the fundamental unit of chromatin. The modification in the structure of chromatin to enable DNA-protein interaction for the processes like replication, transcription is referred to as chromatin remodeling.
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Mammals contain a diploid genome consisting of at least 109 bp. If this amount of DNA is present as chromatin fibers, where each group of 200 bp of DNA is combined with 9 histones into a nucleosome and each group of 6 nucleosomes is combined into a solenoid, achieving a final packing ratio of 50, determine (a) the total number of nucleosomes in all fibers, (b) the total number of histone molecules combined with DNA in the diploid genome, and (c) the combined length of all fibers.
You would like to add a nuclear localization sequence (NLS) of Lys-Lys-Lys-Arg-Lys to a
protein that is usually found in the cytoplasm of a yeast cell. To accomplish this, you
introduce the nucleotide sequence encoding the NLS into the gene that encodes the
cytoplasmic protein of interest.
a. What is the size of the nucleotide insert that will encode the NLS? Briefly explain.
5'
3'
b. Below is a diagram of the gene encoding the cytoplasmic protein of interest in
the yeast genome. If your goal is to put the NLS at the carboxyl (C) terminus of
the protein, at which location (A-E) should the NLS be inserted? Briefly explain.
A
TATAA
ATATT
promoter
+1
B
ATG
TAC
D
TAA
ATT
stop
codon
E
3'
5'
To hold bacterial DNA in a more compact configuration, specificproteins must bind to the DNA and stabilize its conformation.Several different proteins are involved in this process. Some of theseproteins, such as H-NS, have been referred as histone-like, due totheir functional similarity to the histone proteins found in eukaryotes.Based on your knowledge of eukaryotic histone proteins, whatbiochemical properties would you expect bacterial histone-likeproteins to have?
Chapter 12 Solutions
EBK CONCEPTS OF GENETICS
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- The steroid progesterone has an important role in the female reproductive system. Researchers interested in studying membrane progestin receptors (MPRS) developed a method to produce and purify the protein in active form. First, the researchers devised a way to prepare a specific MPR known as hMPRA using the machinery of yeast cells. In order to facilitate purification and identification in later studies, they manipulated the yeast cells so that they attached two different tags to the C-terminal end of the protein. The first tag, Compound 1, is a peptide sequence that acts as an epitope, part of a much larger peptide sequence that is recognized by the immune system. The second sequence consisted of six consecutive histidine residues (His). This sequence binds tightly to Ni2+ cations. In chromatography, (His), tag labeled proteins can be eluted from Ni²+. supported columns by adding a small molecule to the eluent that mimics the side chain of histidine.…arrow_forwardThe steroid progesterone has an important role in the female reproductive system. Researchers interested in studying membrane progestin receptors (MPRs) developed a method to produce and purify the protein in active form. First, the researchers devised a way to prepare a specific MPR known as hMPRA using the machinery of yeast cells. In order to facilitate purification and identification in later studies, they manipulated the yeast cells so that they attached two different tags to the C-terminal end of the protein. The first tag, Compound 1, is a peptide sequence that acts as an epitope, part of a much larger peptide sequence that is recognized by the immune system. The second sequence consisted of six consecutive histidine residues (His). This sequence binds tightly to Ni2+ cations. In chromatography, (His), tag labeled proteins can be eluted from Ni²+- supported columns by adding a small molecule to the eluent that mimics the side chain of histidine. After preparing hMPRA, the…arrow_forwardIn the first figure of “DNMT3L Connects Unmethylated Lysine 4 of Histone H3 to de novo Methylation of DNA,” the authors determined that the DNMT3L protein interacts with several histone proteins. Using Figure a and b (from that paper), explain how each of the 4 histones interact with DNMT3L.arrow_forward
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