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Bacterial Transformation

Engineering bacteria using DNA plasmid

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Bacterial Transformation

A natural process that occurs in some bacteria like E.coli, where they can take up genetic material (DNA) from their environment and incorporate it into their own genome.

The process is important in genetics research and biotechnology, as it allows scientists to introduce new genes into bacteria for various purposes.

Bacterial transformation is a fundamental process in genetic engineering and biotechnology. It allows scientists to manipulate bacterial DNA and create organisms with specific traits, which can have important applications in medicine, agriculture, and industry.

Some real-world applications include insulin production, genetically modified crops, and vaccine development

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The Process

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Scientists first isolate the DNA containing the gene of interest. This gene could code for a specific trait, like making bacteria glow in the dark or produce its own color pigment

Preparation of DNA

Preparing Bacteria

The bacteria that will be transformed (usually E.coli) are prepared. They’re grown in a lab under controlled conditions to ensure they’re healthy and actively dividing

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Making Bacteria “Competent”

Bacteria are naturally resistant to taking up foreign DNA, so scientists need to make them more receptive to transformation. This is done by treating them with special solutions that make their cell membranes more permeable, allowing the foreign DNA to enter.

Mixing DNA and Bacteria

The prepared DNA and the competent bacteria are mixed. Because the bacterial cell membranes are more permeable, some of the DNA molecules are able to slip through and enter the bacterial cells.

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The Process

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Once inside the bacterial cells, the foreign DNA can be incorporated into the bacterial genome through a process called recombination. The bacterial cell's machinery integrates the new DNA into its own DNA sequence.

Incorporation of DNA

Selection

Not all bacteria will successfully take up the new DNA. To identify the ones that have undergone transformation, scientists use a selection process. For instance, if the foreign DNA carries a gene for antibiotic resistance, the transformed bacteria can be grown in an environment containing that antibiotic. Only the transformed bacteria will survive, as they now possess the antibiotic resistance gene.

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Verification

Scientists further verify the presence of the desired DNA in the transformed bacteria using techniques like PCR (polymerase chain reaction) or gel electrophoresis.

Application

Once the transformed bacteria have been identified and confirmed, they can be used for various purposes. For example, they can be used to produce useful proteins, study gene functions, or even create genetically modified organisms.

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Engineer-it Kit! �Transforming K12 E.coli cells with a DNA plasmid

Bacterial transformation is an important technique that has many practical applications in today’s world, such as medicine production, genetic research, crop improvement, etc. At Amino Labs we have a hands-on approach to learning genetic engineering using the Engineer-it Kit. By the end of this project, you’ll have grown and engineered bacteria to produce a colorful bio-pigment!

In the Engineer-it Kit, you will insert circular strands of DNA, called DNA plasmids, into cells. The DNA plasmids contain genes that have been designed to make your cells resistant to common laboratory antibiotic and to produce a coloured pigment.

You will also explore how we can make bacteria competent. To make chemically competent cells, we mix ‘blank cells’ into cold transformation buffer (T. Buffer) and creating cells immediately ready for transformation.

To get the DNA into the cells, you must heatshock the cells. During the shock, the DNA enters the cells.

Since the heatshock step was harmful to the cells, you must ‘recover’ them �using the Recovery media.

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Three Steps to Microfacturing

Let’s look at how DNA can be ‘recognized,’ ‘read,’ and ‘executed’ by the cell.

Step 1: Transcription

Step 2: Translation

Step 3: Enzymatic Processing

DNA becomes bound to and is ‘read’ by cellular machinery called RNA polymerase. RNA polymerase travels along the DNA reading it and simultaneously creates the appropriate RNA molecule.

RNA is less stable than DNA but has similar traits. Ribosomes, along with transfer RNA, recognize an RNA region through bonding. They work together to read and translate the RNA sequence into protein-building blocks called amino acids.

Protein is used to catalyze a chemical reaction involving other molecules which leads to the desired product(s).

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JEOPARDY BOARD

Bacterial Transformation	The Process	Engineer-it Kit	Microfacturing	Terminology
$100	$100	$100	$100	$100
$200	$200	$200	$200	$200
$300	$300	$300	$300	$300
$400	$400	$400	$400	$400
$500	$500	$500	$500	$500

FINAL JEOPARDY

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Bacterial Transformation - $100 Question

The natural process in bacteria where they absorb DNA from their surroundings and integrate it into their own genetic material.

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