3) (3 points) The purpose of the next question is for you to develop an understa

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3) (3 points) The purpose of the next question is for you to develop an understanding of the various experimental steps that are required to do recombinant DNA work. For this question, create numbered list of experimental steps, like a protocol. Use class notes, the book, online material, and other references to find the details of the procedures (most details will be in class notes and book). Specific sequences, compositions of buffers, and complete ingredient lists arc not needed, but include key reagents needed to carry out the specific steps. I have a hypothesis that a certain growth factor receptor x (we?ll call it GFRX) becomes activated in brain tumors and I want to study this receptor to understand it better. Starting with a sample of brain tissue, make a cDNA library (collection of bacteria containing cDNA of all expressed genes in tissue).

Explanation / Answer

The brain tissue sample is collected. The DNA is extracted. The gene encoding the receptor of growth factor X is identified (target gene). This can be identified by DNA sequencing or micro array or RFLP techniques. A sitable bacterium is selected. The plasmid DNA and target DNA are cut with same restriction endonuclease. The gene is to be inserted and cloned in plasmid DNA. The transformed gene is screened and detected in plasmid DNA. This bacteria is incubated with the brain tissue and the gene action in the tissue is studied. The receptor properties are recorded by careful observation of bacteria with target gene interacting with the brain tissue.

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