Suppose you have a 1-kb segment of cloned DNA that is suspected to contain a eukaryotic promoter including a TATA box, a CAT box, and an upstream GC-rich sequence. The clone also contains a gene whose transcript is readily detectable. Your laboratory supervisor asks you to outline an experiment. (1) determine if eukaryotic transcription factors (TF) bind to the fragment and, if so, (2) identify where on the fragment the transcription factors bind.
Gene Interactions
When the expression of a single trait is influenced by two or more different non-allelic genes, it is termed as genetic interaction. According to Mendel's law of inheritance, each gene functions in its own way and does not depend on the function of another gene, i.e., a single gene controls each of seven characteristics considered, but the complex contribution of many different genes determine many traits of an organism.
Gene Expression
Gene expression is a process by which the instructions present in deoxyribonucleic acid (DNA) are converted into useful molecules such as proteins, and functional messenger ribonucleic (mRNA) molecules in the case of non-protein-coding genes.
Suppose you have a 1-kb segment of cloned DNA that is suspected to contain a eukaryotic promoter including a TATA box, a CAT box, and an upstream GC-rich sequence. The clone also contains a gene whose transcript is readily detectable. Your laboratory supervisor asks you to outline an experiment. (1) determine if eukaryotic transcription factors (TF) bind to the fragment and, if so, (2) identify where on the fragment the transcription factors bind. All necessary reagents, equipment, and experimental know-how are available in the laboratory.
Complete the outline of an experiment, which determines if eukaryotic transcription factors (TF) bind to the fragment. Assume all necessary reagents, equipment, and experimental how are available in the laboratory.
Drag the terms on the left to the appropriate blanks on the right to complete the sentences. Not all terms will be used.
For the band shift assay, two samples of the DNA fragment are analyzed by agarose gel electrophoresis: one sample is mixed with TFs ( ), and one is treated identically except without the TFs ( ). After allowing sufficient time for TFs to bind to DNA, the samples are subjected to gel electrophoresis and the relative mobility of the two DNA samples is compared. If TFs are __________ and ________ to the DNA, the experimental sample will have a __________ electrophoretic mobility than the control sample.
Answer :- The goal of this experiment is to determine if eukaryotic transcription factors (TFs) bind to a specific 1-kilobase (kb) segment of cloned DNA suspected to contain a eukaryotic promoter region, including a TATA box, a CAT box, and an upstream GC-rich sequence. Additionally, we aim to identify where on the DNA fragment these transcription factors might bind. The presence of TF binding is crucial as it indicates the regulatory potential of this DNA segment.
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