You have identified a recessive mutation that alters bristle patterning in Drosophila and have used recombinant DNA technology to identify a genomic clone that you believe harbors the gene. How would you demonstrate that your gene is on the genomic clone?

Verified step by step guidance

1. Step 1: Perform a complementation test by introducing the genomic clone into a Drosophila strain that is homozygous for the recessive mutation. Observe if the wild-type bristle pattern is restored, indicating that the clone contains the functional gene.
2. Step 2: Conduct a rescue experiment by inserting the genomic clone into a vector suitable for Drosophila transformation and introduce it into mutant flies. Check if the bristle pattern is corrected in the transformed flies.
3. Step 3: Use Southern blot analysis to confirm the presence of the genomic clone in the transformed flies' DNA, ensuring that the clone has integrated into the genome.
4. Step 4: Perform a sequence analysis of the genomic clone to identify the gene of interest and compare it with known sequences to confirm its identity.
5. Step 5: Use RNA in situ hybridization or RT-PCR to verify that the gene on the genomic clone is expressed in tissues where bristle patterning occurs, supporting its role in the phenotype.

Key Concepts

Recessive Mutation

A recessive mutation is a genetic alteration that only manifests phenotypically when an individual has two copies of the mutated allele. In the context of Drosophila, this means that the altered bristle patterning will only be observed in flies that inherit the recessive allele from both parents. Understanding this concept is crucial for determining how the mutation affects the phenotype and for designing experiments to identify the responsible gene.

Recombinant DNA Technology

Recombinant DNA technology involves combining DNA from different sources to study specific genes or genetic functions. This technique allows researchers to isolate and clone genes of interest, such as the one responsible for the bristle patterning mutation in Drosophila. By using this technology, you can create genomic clones that may contain the gene, facilitating further analysis and experimentation to confirm its role in the observed phenotype.

Genomic Cloning and Gene Mapping

Genomic cloning is the process of isolating and amplifying a specific segment of DNA, which can then be analyzed to determine its function. Gene mapping involves identifying the location of a gene within a genome, often using techniques like PCR, sequencing, or hybridization. To demonstrate that the gene is on the genomic clone, you would typically perform experiments such as complementation tests or gene expression analysis to show that the cloned DNA can restore the wild-type phenotype in the presence of the recessive mutation.