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Prokaryote reproduction and biotechnology
How prokaryotes reproduce by binary fission. Use of E. coli bacteria in molecular biology.
Key points:
- Prokaryotes (bacteria and archaea) reproduce asexually through binary fission. Most prokaryotes reproduce rapidly.
- Due to their fast growth and simple genetics, E. coli bacteria are widely used in molecular biology.
- In the laboratory, a gene can be transferred into E. coli bacteria on a small, circular DNA molecule called a plasmid. The plasmid is taken up by the bacteria in a process called transformation.
- The transformed E. coli bacteria can be used to make many copies of the plasmid. In some cases, they will also express the gene on the plasmid and make protein.
Introduction
Let's say you have one bacterium. How can you get more identical bacteria? How quickly can you get them? And, most importantly, why on Earth would you want a whole bunch of identical bacteria?
Let's fast-forward to that last question: some bacteria, most notably Escherichia coli (E.coli), are widely used in molecular biology labs. There, they serve as little "factories" that churn out many copies of a desired DNA molecule, or many molecules of a needed protein (such as the insulin used by diabetics to regulate their blood sugar). The more bacteria, the more of the DNA or protein product that can be made.
Two features that make E. coli very useful in the lab are its rapid reproduction and its generation of clones, or genetically identical bacteria. Let's take a quick look at how E. coli and other prokaryotes reproduce. Then, we'll examine their applications in biotechnology.
How do prokaryotes reproduce?
Prokaryotes reproduce through a cell division process called binary fission. Like mitosis in eukaryotes, this process involves copying the chromosome and separating one cell into two.
Binary fission is an asexual form of reproduction, meaning that it does not involve production of eggs and sperm or mixing of genetic material from two individuals. Except in the case of rare mutations, or changes in DNA sequence, binary fission produces daughter cells that are genetically identical to the mother cell.
You can learn more about the steps of binary fission in the binary fission article in the cell division section.
Prokaryotes reproduce fast!
Prokaryotes in general reproduce much faster than multicellular eukaryotes. This can be measured in terms of generation time, or the length of time from the birth of one generation to the birth of the next.
For humans, a typical generation time might be in the neighborhood of
Not all bacteria are quite this quick, and some pathogenic ones, such as Mycobacterium tuberculosis, have a generation time over
Bacteria in molecular biology
Bacteria that reproduce quickly and are easy to grow in the lab make good model organisms for many scientific studies. E. coli, for instance, is one of the most widely used organisms in biological research.
Although you may have heard of E. coli as a food contaminant, harmless strains of E. coli are used in biology labs worldwide. In fact, many basic biological processes, like the mechanism of DNA replication, were first discovered in E. coli.
E. coli as DNA and protein factories
Today, E. coli are sometimes used as tiny “factories” to synthesize DNA or proteins. Researchers can insert a gene of interest into E. coli cells through a process called transformation (uptake of DNA from the environment), which is described further in the article on prokaryote genetic variation. In such experiments, the gene of interest is typically borne on a piece of circular DNA called a plasmid, which can be copied by the bacterium and passed on to its offspring.
Once they contain the plasmid with the gene of interest, the E. coli cells will replicate it and pass it along each time they divide, making many copies of the plasmid DNA. If the plasmid contains the right control sequences, the E. coli can also be instructed to transcribe and translate the gene of interest, producing protein. For example, most of the insulin used by diabetics is produced in E. coli cells using this strategy.
Steps of transformation
In a typical transformation experiment, the target gene (blue DNA above) is first inserted into a plasmid. In addition to the target gene, the plasmid also contains a gene that provides resistance to a particular antibiotic (red DNA above). If the goal is to use the bacteria to synthesize protein from the gene, the plasmid will also contain a promoter, or control sequence, that allows the target gene to be expressed in bacteria (green DNA above).
When copies of the plasmid are mixed with E. coli cells and the cells are heat-shocked (exposed briefly to high temperature), a small fraction of them will take up the plasmid. All of the E. coli are then spread on a nutrient plate containing the antibiotic. The purpose of the antibiotic is to only let bacteria with the plasmid survive and grow.
E. coli lacking the plasmid will be killed by the antibiotic. E. coli that contain the plasmid, however, can survive and reproduce (thanks to the antibiotic resistance gene in the plasmid). Each resistant cell will form a colony of genetically identical bacteria, which appears on the agar plate as small dot. An antibiotic-resistant colony can be analyzed (checked by other methods to confirm it contains the correct plasmid), then grown up to make a large culture of identical, plasmid-bearing bacteria.
What use is a large culture of plasmid-bearing bacteria? Sometimes, researchers need many copies of the plasmid DNA for use in another experiment, and they can extract this DNA from the culture. Alternatively, if the plasmid contains the right promoter, the bacteria can be induced (instructed) to express the gene and synthesize protein. This technique is used to produce some medically important proteins, such as insulin and human growth hormone.
Want to join the conversation?
what are gram positive and negative bacteria(4 votes)
Gram-positive bacteria are the ones which, when a stain is added (in the Gram staining technique), become purple. Gram-negative bacteria lose the purple stain easily, and they only retain a red or pink color. This is because gram-positive bacteria have a thick peptidoglycan (sugar and amino acid) layer which absorbs the purple stain very well, but gram-negative ones have a very thin peptidoglycan layer, so they are only weakly stained.(14 votes)
Has CRISPR largely replaced the method of transformation explained in this article?(3 votes)
CRISPR followed by recombination can be an alternative to restriction followed by ligation.
It can't replace transforming DNA into bacteria.
In fact, we generally assemble the parts needed to make CRISPR work in a bacterial plasmid!(3 votes)
- Why does a heat shock trigger the bacterium to take the plasmid?
How does a bacterium take up the plasmid?
And what are some possible causes of bacteria not taking them?(2 votes)- Heat shock increases the permeability of the cell membrane so that the plasmids can move across it more easily, the bacteria will not immediately take up the DNA so the cell membrane has to be destabilized by the heat shock so that the plasmids move across it. This is not a perfect process so only some bacteria are transformed (take up the plasmid) because some of it is due to chance. The bacteria do not really choose whether or not to take the plasmid, it is more just if the cell membrane is permeable to the plasmid or if it is not.(4 votes)
Whats the difference between eukaryotic reproduction and prokaryotic reproduction?(2 votes)
The main difference is that mitosis requires microtubules to pull the chromosomes apart, whereas binary fission just needs the nucleoid to replicate then the bacteria can immediately divide.(3 votes)
can PCR amplify prokaryotic cells?(2 votes)- It does not amplify cells but isolated DNA, it does not matter where you take the DNA from.(3 votes)
Are plasmids something that humans create and they give them to bacteria? From the article above, it sounds like humans create and give them to bacteria.(2 votes)
No, humans do not make plasmid DNA. All human DNA is found within our linear chromosomes. Plasmids are generally found only in bacteria (with the exception of some archaea and eukaryota). The plasmids can come from the circular bacterial genome - bits of DNA can be turned into plasmids and conversely plasmids can be integrated back into the primary genome. Plasmids may also have viral origins, meaning they are a relic of something that infected the bacteria in the past. The system is very fluid, allowing bacteria to share important genetic material easily and fast.(3 votes)
Would E. coli be a gram negative bacteria or gram positive??(2 votes)
It is gram negative, catalase positive (reacts with H2O2), MacConckey agar positive (ferments lactose), and Quellung positive.(2 votes)
What is negative bacteria?(2 votes)- Gram-negative bacteria are bacteria that do not retain the crystal violet stain used in the Gram staining method of bacterial differentiation. So they color as red after Gram staining. E.g. E.coli.(2 votes)
What does Plasmid do in cells? And why do we need Plasmid?(2 votes)- We do not need plasmid.
Plasmids are naturally occurring in Bacteria and rarely in Eukaryotic cells.
For Bacteria, which have nucleoid, the plasmid is pretty useful carrying additional genes such as antibiotic resistance or lethal mutation. It also helps horizontal gene transfer. And not to mention Scientific studies where we can artificially introduce wanted molecule via plasmid into target cell.(2 votes)
- Could this in theory, mean that the insulin produced carry a gene thats resistant to a particular antibiotic? And could that have effects on a diabetic individual using that? (hope this makes sense!)(2 votes)
- Insulin is not a type of organism but hormone our body produces and uses. It does not carry genetic material or whatsoever especially has nothing to do with antibiotics.(2 votes)