You are on a mission to mass-produce the \"Elasticity\" protein from Mr. Fantast

Question

You are on a mission to mass-produce the "Elasticity" protein from Mr. Fantastic. A Marvel agent of S.H.I.E.L.D. has provided you with clean, uncontaminated DNA. Do a DETAILED DIAGRAM INDICATING EACH STEP you will follow from amplification of the gene to verifying that your end product doesn't have an unexpected mutation. You must use proper names for each technique. Think about controls. (Steps: amplifying gene of interest (PCR) and electrophoresis, cleaving DNA, ligation, transformation, screening, DNA purification, and sequencing.

Explanation / Answer

The mission is to produce Elasticity protein from a clean, uncontaminated DNA . DNA has to undergo different steps like DNA cleavage, cloning by PCR, separation by electrophoresis, sequencing, ligation, transformation, replication, transcription and translation finally to synthesize a protein.

Cleavage of DNA at specific sites by restriction nucleases, which greatly facilitates the isolation and manipulation of individual genes.

Separation and Analysis of macromolecules and their fragments like DNA(charged) based on their size and charge can be performed by Electrophoresis.

DNA cloning either through the use of cloning vectors or the polymerase chain reaction, whereby a single DNA molecule can be copied to generate many billions of identical molecules.

Nucleic acid hybridization, which makes it possible to find a specific sequence of DNA or RNA with great accuracy and sensitivity on the basis of its ability to bind a complementary nucleic acid sequence.

Rapid sequencing of all the nucleotides in a purified DNA fragment, which makes it possible to identify genes and to deduce the amino acid sequence of the proteins they encode.

Simultaneous monitoring of the expression level of each gene in a cell, using nucleic acid microarrays that allow tens of thousands of hybridization reactions to be performed simultaneously.

Ligation of DNA inserts into an appropriate cloning vector, creating recombinant molecules (e.g., plasmids)

Transformation of recombinant plasmids into bacteria or other suitable host for propagation.

Screening/selection of hosts containing the intended recombinant plasmid.

Replication DNA Replication is the process of making 2 identical copies of DNA from one original DNA copy.

Transcription of template DNA strand occurs where it is converted to complementary coding RNA (m RNA) to begin gene expression.

Translation Translation is the process by which a RNA sequence is converted into a set of amino acids ( a protein). Each amino acid is coded for by a 3 nucleotide sequence called a codon.

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