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Ch. 8 - Microbial Genetics
Chapter 8, Problem 9.6a

Describe an rDNA experiment in two or three sentences. Use the following terms: intron, exon, DNA, mRNA, cDNA, RNA polymerase, reverse transcriptase.

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Start by isolating the mRNA from a eukaryotic cell, which contains both exons and introns.
Use reverse transcriptase to synthesize complementary DNA (cDNA) from the mRNA template, which will only include exons since introns are removed during mRNA processing.
The cDNA is then inserted into a plasmid vector to create recombinant DNA (rDNA).
Introduce the plasmid into a bacterial host cell, where RNA polymerase will transcribe the cDNA into mRNA, allowing for protein expression.
Finally, analyze the expression of the protein to confirm the success of the rDNA experiment.

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Key Concepts

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

Recombinant DNA (rDNA) Technology

Recombinant DNA technology involves combining DNA from different sources to create new genetic combinations. This process often includes the use of enzymes to cut and join DNA fragments, allowing scientists to manipulate genes for various applications, such as gene cloning or the production of proteins. Understanding rDNA is crucial for designing experiments that involve gene expression and manipulation.
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Map of DNA-Based Technology Lesson

Transcription and Reverse Transcription

Transcription is the process by which RNA polymerase synthesizes mRNA from a DNA template, transcribing the exons (coding regions) while excluding introns (non-coding regions). In contrast, reverse transcription is the process where reverse transcriptase converts mRNA back into cDNA (complementary DNA). This cDNA can then be used in various applications, including cloning and gene expression analysis.
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Initiation of Transcription

mRNA and cDNA

mRNA (messenger RNA) is the product of transcription and carries genetic information from DNA to the ribosome for protein synthesis. cDNA (complementary DNA) is synthesized from mRNA using reverse transcriptase and serves as a stable form of the gene that can be cloned or analyzed. Understanding the relationship between mRNA and cDNA is essential for experiments involving gene expression and the study of specific genes.
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Monocistronic mRNA vs. Polycistronic mRNA