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Ch. 15 - Recombinant DNA Technology and Its Applications
Sanders - Genetic Analysis: An Integrated Approach 3rd Edition
Sanders3rd EditionGenetic Analysis: An Integrated ApproachISBN: 9780135564172당신이 사용하는 게 아니라요?교과서 변경
15장, 문제 26b

In Drosophila, loss-of-function Ultrabithorax mutations result in the posterior thoracic segments differentiating into body parts with an identity normally found in the anterior thoracic segments. When the Ultrabithorax gene was cloned, it was shown to encode a transcription factor and to be expressed only in the posterior region of the thorax. Thus, Ultrabithorax acts to specify the identity of the posterior thoracic segments. Similar genes were soon discovered in other animals, including mice and humans. You have found that mice possess two closely related genes, Hoxa7 and Hoxb4, which are orthologs of Ultrabithorax. You wish to know whether the two mouse genes act to specify the identity of body segments in mice.
How will you create loss-of-function alleles of the mouse genes?

검증된 단계별 안내
1
Step 1: Understand the concept of loss-of-function alleles. These are mutations that result in the reduction or complete loss of the normal function of a gene. In this case, you aim to create loss-of-function alleles for the mouse genes Hoxa7 and Hoxb4.
Step 2: Use gene-targeting techniques to create loss-of-function alleles. One common method is homologous recombination in embryonic stem (ES) cells. Design a targeting vector containing a selectable marker (e.g., neomycin resistance gene) flanked by sequences homologous to the regions surrounding the Hoxa7 or Hoxb4 gene.
Step 3: Introduce the targeting vector into mouse ES cells. The homologous recombination process will replace the functional gene with the disrupted version containing the selectable marker, effectively creating a loss-of-function allele.
Step 4: Select and screen ES cells for successful recombination events. Use PCR or Southern blotting to confirm that the targeted gene has been disrupted in the ES cells.
Step 5: Inject the modified ES cells into mouse blastocysts to create chimeric mice. Breed these chimeric mice to produce offspring that are homozygous for the loss-of-function alleles of Hoxa7 and Hoxb4. Analyze the phenotypes of these mice to determine the role of these genes in specifying body segment identity.

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Loss-of-Function Mutations

Loss-of-function mutations are genetic alterations that result in the reduced or abolished activity of a gene product, typically a protein. In the context of the Ultrabithorax gene in Drosophila, such mutations lead to the transformation of posterior thoracic segments into anterior-like structures. Understanding how to create these mutations in mouse genes, like Hoxa7 and Hoxb4, is crucial for studying their roles in segment identity.
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가이드 코스
08:26
Functional Genomics

Transcription Factors

Transcription factors are proteins that bind to specific DNA sequences to regulate the transcription of genes. The Ultrabithorax gene encodes a transcription factor that is essential for specifying the identity of thoracic segments in Drosophila. By understanding the function of transcription factors, researchers can infer how similar genes in mice might influence body segment identity and development.
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가이드 코스
09:16
Eukaryotic Transcription

Orthologs

Orthologs are genes in different species that evolved from a common ancestral gene and typically retain similar functions. The Hoxa7 and Hoxb4 genes in mice are orthologs of the Ultrabithorax gene in Drosophila, suggesting that they may have conserved roles in segment identity. Recognizing the significance of orthologs helps in predicting the function of genes across species and designing experiments to test their roles.
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교과서 질문

In Drosophila, loss-of-function Ultrabithorax mutations result in the posterior thoracic segments differentiating into body parts with an identity normally found in the anterior thoracic segments. When the Ultrabithorax gene was cloned, it was shown to encode a transcription factor and to be expressed only in the posterior region of the thorax. Thus, Ultrabithorax acts to specify the identity of the posterior thoracic segments. Similar genes were soon discovered in other animals, including mice and humans. You have found that mice possess two closely related genes, Hoxa7 and Hoxb4, which are orthologs of Ultrabithorax. You wish to know whether the two mouse genes act to specify the identity of body segments in mice.

How will you determine where and when the mouse genes are expressed?

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교과서 질문

Bacterial Pseudomonas species often possess plasmids encoding genes involved in the catabolism of organic compounds. You have discovered a strain that can metabolize crude oil and wish to identify the gene(s) responsible. Outline an experimental protocol to find the gene or genes required for crude oil metabolism.

515
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교과서 질문

Two complaints about some transgenic plants presently in commercial use are that (1) the Bt toxin gene is constitutively expressed in them, leading to fears that selection pressures will cause insects to evolve resistance to the toxin, and (2) a selectable marker gene—for example, conferring kanamycin resistance—remains in the plant, leading to concerns about increased antibiotic resistance in organisms in the wild. How would you generate transgenic plants that produce Bt only in response to being fed upon by insects and without the selectable marker?

428
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교과서 질문

You have identified an enhancer trap line generated by P element transposition in Drosophila in which the marker gene from the enhancer trap is specifically expressed in the wing imaginal disc.

How can you identify the gene adjacent to the insertion site of the enhancer trap?

573
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교과서 질문

You have identified an enhancer trap line generated by P element transposition in Drosophila in which the marker gene from the enhancer trap is specifically expressed in the wing imaginal disc.

How would you show that the expression pattern of the enhancer trap line reflects the endogenous gene expression pattern of the adjacent gene?

572
views
교과서 질문

In Drosophila, loss-of-function Ultrabithorax mutations result in the posterior thoracic segments differentiating into body parts with an identity normally found in the anterior thoracic segments. When the Ultrabithorax gene was cloned, it was shown to encode a transcription factor and to be expressed only in the posterior region of the thorax. Thus, Ultrabithorax acts to specify the identity of the posterior thoracic segments. Similar genes were soon discovered in other animals, including mice and humans. You have found that mice possess two closely related genes, Hoxa7 and Hoxb4, which are orthologs of Ultrabithorax. You wish to know whether the two mouse genes act to specify the identity of body segments in mice.

How will you determine whether the mouse genes have redundant functions?

544
views