Table of contents

1. Introduction to Genetics51m
2. Mendel's Laws of Inheritance3h 37m
3. Extensions to Mendelian Inheritance2h 41m
4. Genetic Mapping and Linkage2h 28m
5. Genetics of Bacteria and Viruses1h 21m
6. Chromosomal Variation1h 48m
7. DNA and Chromosome Structure56m
8. DNA Replication1h 10m
9. Mitosis and Meiosis1h 34m
10. Transcription1h 0m
11. Translation58m
12. Gene Regulation in Prokaryotes1h 19m
13. Gene Regulation in Eukaryotes44m
14. Genetic Control of Development44m
15. Genomes and Genomics1h 50m
16. Transposable Elements47m
17. Mutation, Repair, and Recombination1h 6m
18. Molecular Genetic Tools19m
- Genetic Cloning
  9m
- Methods for Analyzing DNA
  9m
19. Cancer Genetics29m
- Overview of Cancer
  21m
- Cancer Mutations
  7m
20. Quantitative Genetics1h 26m
21. Population Genetics50m
- Hardy Weinberg
  19m
- Allelic Frequency Changes
  31m
22. Evolutionary Genetics29m

10. Transcription

Transcription in Eukaryotes

Genetics

10. Transcription

Transcription in Eukaryotes

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2:04 minutes

Problem 15

Textbook Question

Many promoter regions contain CAAT boxes containing consensus sequences CAAT or CCAAT approximately 70 to 80 bases upstream from the transcription start site. How might one determine the influence of CAAT boxes on the transcription rate of a given gene?

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Understand the role of CAAT boxes: CAAT boxes are regulatory elements in the promoter region of a gene that influence the binding of transcription factors and RNA polymerase, thereby affecting the transcription rate.

Design an experiment to mutate the CAAT box: Use site-directed mutagenesis to alter the CAAT or CCAAT sequence in the promoter region of the gene. This will help determine the effect of the CAAT box on transcription.

Create reporter gene constructs: Clone the wild-type promoter (with the intact CAAT box) and the mutated promoter (with the altered CAAT box) upstream of a reporter gene, such as luciferase or GFP, in separate plasmids.

Transfect cells with the constructs: Introduce the plasmids into a suitable cell line using a transfection method, such as electroporation or lipofection, to allow the cells to express the reporter gene under the control of the respective promoters.

Measure transcriptional activity: Quantify the reporter gene expression (e.g., by measuring luciferase activity or GFP fluorescence) for both the wild-type and mutated constructs. Compare the results to determine the influence of the CAAT box on transcription rate.

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Key Concepts

Here are the essential concepts you must grasp in order to answer the question correctly.

Promoter Regions

Promoter regions are specific DNA sequences located upstream of a gene that serve as binding sites for RNA polymerase and transcription factors. They play a crucial role in initiating transcription, as they determine when and how much of a gene is expressed. The presence of specific elements, such as CAAT boxes, can enhance the binding of transcription machinery, thereby influencing the transcription rate.

CAAT Boxes

CAAT boxes are conserved DNA sequences found in the promoter regions of many eukaryotic genes, typically located 70 to 80 base pairs upstream of the transcription start site. These sequences are recognized by transcription factors that enhance the efficiency of transcription initiation. The presence and integrity of CAAT boxes can significantly affect the transcriptional activity of associated genes, making them important for gene regulation.

Transcription Rate

The transcription rate refers to the speed at which RNA polymerase synthesizes RNA from a DNA template during the transcription process. This rate can be influenced by various factors, including the presence of promoter elements like CAAT boxes, the availability of transcription factors, and the overall chromatin structure. Understanding how these elements interact helps in determining the expression levels of specific genes.

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Compare the control of gene regulation in eukaryotes and bacteria at the level of initiation of transcription. How do the regulatory mechanisms work? What are the similarities and differences in these two types of organisms in terms of the specific components of the regulatory mechanisms?

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The eukaryotic gene Gen-100 contains four introns labeled A to D. Imagine that Gen-100 has been isolated and its DNA has been denatured and mixed with polyadenylated mRNA from the gene.

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