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Ch. 6 - Genetic Analysis and Mapping in Bacteria and Bacteriophages
Sanders - Genetic Analysis: An Integrated Approach 3rd Edition
Sanders3rd EditionGenetic Analysis: An Integrated ApproachISBN: 9780135564172Not the one you use?Change textbook
Chapter 6, Problem 18a

An interrupted mating study is carried out on Hfr strains 1, 2, and 3 identified in Problem 17. After conjugation is established, a small sample of the mixture is collected every minute for 20 minutes to determine the distance between genes on the chromosome. Results for each of the three Hfr strains are shown below. The total duration of conjugation (in minutes) is given for each transferred gene.
Hfr strain 1     oriT  met  ala  lac  gal
Duration (min)   0     2    8   13   17
Hfr strain 2     oriT  met  leu  thr   azi
Duration (min)   0     2    7   10   17
Hfr strain 3     oriT  gal   pro  trp   azi
Duration (min)   0     3    8   14   19
For each Hfr strain, draw a time-of-entry profile.

Verified step by step guidance
1
Step 1: Understand the concept of an interrupted mating study. This technique is used to map the relative positions of genes on a bacterial chromosome by determining the time at which specific genes are transferred during conjugation. The 'oriT' (origin of transfer) is the starting point for gene transfer, and the duration indicates the time it takes for each gene to be transferred.
Step 2: For each Hfr strain, plot a time-of-entry profile. On the x-axis, represent the duration of conjugation in minutes (0 to 20 minutes). On the y-axis, list the genes in the order they are transferred. For example, for Hfr strain 1, the genes are oriT, met, ala, lac, and gal.
Step 3: Mark the time points on the x-axis corresponding to the transfer duration for each gene. For Hfr strain 1, plot oriT at 0 minutes, met at 2 minutes, ala at 8 minutes, lac at 13 minutes, and gal at 17 minutes. Repeat this process for Hfr strains 2 and 3 using their respective durations and genes.
Step 4: Connect the points for each strain with a line to create a time-of-entry profile. This line visually represents the order and timing of gene transfer during conjugation for each Hfr strain.
Step 5: Compare the profiles of the three Hfr strains to identify differences in gene order and transfer timing. This comparison helps determine the relative positions of genes on the bacterial chromosome and the orientation of the 'oriT' in each strain.

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Key Concepts

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

Hfr Strains and Conjugation

Hfr (high frequency of recombination) strains are bacterial strains that have integrated a plasmid into their chromosome, allowing them to transfer chromosomal genes to a recipient bacterium during conjugation. This process involves the formation of a pilus that connects two bacteria, enabling the transfer of genetic material. Understanding Hfr strains is crucial for studying gene mapping and the genetic exchange between bacteria.
Recommended video:
Guided course
04:06
Conjugation Overview

Gene Mapping and Time-of-Entry

Gene mapping involves determining the relative positions of genes on a chromosome based on the frequency of their co-transfer during conjugation. The time-of-entry profile illustrates the sequence and timing of gene transfer, where earlier genes are closer to the origin of transfer (oriT). This method allows researchers to estimate the distances between genes, providing insights into the organization of bacterial genomes.
Recommended video:
Guided course
09:09
Mapping Genes

Interrupted Mating Technique

The interrupted mating technique is a method used to study gene transfer in bacteria by stopping the conjugation process at specific time intervals. By collecting samples at regular intervals, researchers can analyze which genes have been transferred and their order of entry. This technique is essential for constructing genetic maps and understanding the dynamics of bacterial conjugation.
Recommended video:
Guided course
07:55
Non-Random Mating
Related Practice
Textbook Question

Hfr strains that differ in integrated F factor orientation and site of integration are used to construct consolidated bacterial chromosome maps. The data below show the order of gene transfer for five strains.

Hfr Strain   Order of Gene Transfer (First → Last)

Hfr A        oriT–thr–leu–azi–ton–pro–lac–ade

Hfr B        oriT–mtl–xyl–mal–str–his

Hfr C        oriT–ile–met–thi–thr–leu–azi–ton

Hfr D        oriT–his–trp–gal–ade–lac–pro–ton

Hfr E        oriT–thi–met–ile–mtl–xyl–mal–str

Identify the overlaps between Hfr strains. Identify the orientations of integrated F factors relative to one another.

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Textbook Question

Hfr strains that differ in integrated F factor orientation and site of integration are used to construct consolidated bacterial chromosome maps. The data below show the order of gene transfer for five strains.

Hfr Strain   Order of Gene Transfer (First → Last)

Hfr A        oriT–thr–leu–azi–ton–pro–lac–ade

Hfr B        oriT–mtl–xyl–mal–str–his

Hfr C        oriT–ile–met–thi–thr–leu–azi–ton

Hfr D        oriT–his–trp–gal–ade–lac–pro–ton

Hfr E        oriT–thi–met–ile–mtl–xyl–mal–str

Draw a consolidated map of the bacterial chromosome. (Hint: Begin by placing the insertion site for Hfr A at the 2 o'clock position and arranging the genes thr–leu–azi- . . . in clockwise order.)

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Textbook Question

Five Hfr strains from the same bacterial species are analyzed for their ability to transfer genes to F⁻ recipient bacteria. The data shown below list the origin of transfer (oriT) for each strain and give the order of genes, with the first gene on the left and the last gene on the right. Use the data to construct a circular map of the bacterium.

Hfr Strain     Genes Transferred

Hfr 1         oriT met ala lac gal

Hfr 2         oriT met leu thr azi

Hfr 3         oriT gal pro trp azi

Hfr 4         oriT leu met ala lac

Hfr 5         oriT trp azi thr leu met

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Textbook Question

An interrupted mating study is carried out on Hfr strains 1, 2, and 3 identified in Problem 17. After conjugation is established, a small sample of the mixture is collected every minute for 20 minutes to determine the distance between genes on the chromosome. Results for each of the three Hfr strains are shown below. The total duration of conjugation (in minutes) is given for each transferred gene.

Hfr strain 1     oriT  met  ala  lac  gal

Duration (min)   0     2    8   13   17

Hfr strain 2     oriT  met  leu  thr   azi

Duration (min)   0     2    7   10   17

Hfr strain 3     oriT  gal   pro  trp   azi

Duration (min)   0     3    8   14   19

Using the chromosome map you prepared in answer to Problem 17, determine the distance in minutes between each gene on the map.

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Textbook Question

An interrupted mating study is carried out on Hfr strains 1, 2, and 3 identified in Problem 17. After conjugation is established, a small sample of the mixture is collected every minute for 20 minutes to determine the distance between genes on the chromosome. Results for each of the three Hfr strains are shown below. The total duration of conjugation (in minutes) is given for each transferred gene.

Hfr strain 1     oriT  met  ala  lac  gal

Duration (min)   0     2    8   13   17

Hfr strain 2     oriT  met  leu  thr   azi

Duration (min)   0     2    7   10   17

Hfr strain 3     oriT  gal   pro  trp   azi

Duration (min)   0     3    8   14   19

Explain why azi is the last gene of strain 2 to transfer in the 20 minutes of conjugation time. How many minutes of conjugation time would be needed to allow the next gene on the map to transfer from Hfr strain 2?

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Textbook Question

An interrupted mating study is carried out on Hfr strains 1, 2, and 3 identified in Problem 17. After conjugation is established, a small sample of the mixture is collected every minute for 20 minutes to determine the distance between genes on the chromosome. Results for each of the three Hfr strains are shown below. The total duration of conjugation (in minutes) is given for each transferred gene.

Hfr strain 1     oriT  met  ala  lac  gal

Duration (min)   0     2    8   13   17

Hfr strain 2     oriT  met  leu  thr   azi

Duration (min)   0     2    7   10   17

Hfr strain 3     oriT  gal   pro  trp   azi

Duration (min)   0     3    8   14   19

Write out the interrupted mating results you would expect after 20 minutes of conjugation for Hfr strains 4 and 5. Use the format shown at the beginning of this problem.

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