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Plasmids occur naturally in bacterial populations (such as Escherichia coli ) and have genes that can contribute favorable traits to the organism, such as antibiotic resistance (the ability to be unaffected by antibiotics). Plasmids have been highly engineered as vectors for molecular cloning and for the subsequent large-scale production of important molecules, such as insulin. A valuable characteristic of plasmid vectors is the ease with which a foreign DNA fragment can be introduced. These plasmid vectors contain many short DNA sequences that can be cut with different commonly available restriction enzymes . Restriction enzymes (also called restriction endonucleases) recognize specific DNA sequences and cut them in a predictable manner; they are naturally produced by bacteria as a defense mechanism against foreign DNA. Many restriction enzymes make staggered cuts in the two strands of DNA, such that the cut ends have a 2- to 4-nucleotide single-stranded overhang. The sequence that is recognized by the restriction enzyme is a four- to eight-nucleotide sequence that is a palindrome. Like with a word palindrome, this means the sequence reads the same forward and backward. In most cases, the sequence reads the same forward on one strand and backward on the complementary strand. When a staggered cut is made in a sequence like this, the overhangs are complementary ( [link] ).

In part A, the figure shows a strand of ladder-like DNA. In part B, the DNA is cut on both strands between the two guanines. In part C, the 2 strands have separated, leaving complementary sticky ends on each with unattached 5' to 3' G, A, T, and C nucleotides.
In this (a) six-nucleotide restriction enzyme recognition site, notice that the sequence of six nucleotides reads the same in the 5' to 3' direction on one strand as it does in the 5' to 3' direction on the complementary strand. This is known as a palindrome. (b) The restriction enzyme makes breaks in the DNA strands, and (c) the cut in the DNA results in “sticky ends”. Another piece of DNA cut on either end by the same restriction enzyme could attach to these sticky ends and be inserted into the gap made by this cut.

Because these overhangs are capable of coming back together by hydrogen bonding with complementary overhangs on a piece of DNA cut with the same restriction enzyme, these are called “sticky ends.” The process of forming hydrogen bonds between complementary sequences on single strands to form double-stranded DNA is called annealing . Addition of an enzyme called DNA ligase, which takes part in DNA replication in cells, permanently joins the DNA fragments when the sticky ends come together. In this way, any DNA fragment can be spliced between the two ends of a plasmid DNA that has been cut with the same restriction enzyme ( [link] ).

An illustration showing the steps in creating recombinant DNA plasmids, inserting them into bacteria, and then selecting only the bacteria that have successfully taken up the recombinant plasmid. The steps are as follows: both foreign DNA and a plasmid are cut with the same restriction enzyme. The restriction site occurs only once in the plasmid in the middle of a gene for an enzyme (lacZ). The restriction enzyme leaves complementary sticky ends on the foreign DNA fragment and the plasmid. This allows the foreign DNA to be inserted into the plasmid when the sticky ends anneal. Adding DNA ligase reattaches the DNA backbones. These are recombinant plasmids. The plasmids are combined with a culture of living bacteria. Many of the bacteria do not take any plasmids into their cells, many take plasmids that do not have the foreign DNA in them, and a few take up the recombinant plasmid. The bacteria that take up the recombinant plasmid cannot make the enzyme from the gene that the fragment was inserted into (lacZ). They also carry a gene for resistance to the antibiotic ampicillin, which was on the original plasmid. To find the bacteria with the recombinant plasmid, the bacteria are grown on a plate with the antibiotic ampicillin and a substance that changes color when exposed to the enzyme produced by the lacZ gene. The ampicillin will kill any bacteria that did not take up a plasmid. The color of the substance will not change when the gene for lacZ contains the foreign DNA insert. These are the bacteria with the recombinant plasmid that we want to grow.
This diagram shows the steps involved in molecular cloning.

Plasmids with foreign DNA inserted into them are called recombinant DNA    molecules because they contain new combinations of genetic material. Proteins that are produced from recombinant DNA molecules are called recombinant proteins . Not all recombinant plasmids are capable of expressing genes. Plasmids may also be engineered to express proteins only when stimulated by certain environmental factors, so that scientists can control the expression of the recombinant proteins.

Questions & Answers

Cellular respiration
Lucy Reply
what are the characteristics of living things
Ruth Reply
Movement Respiration Nutrition/Feeding Irritability/Sensitivity Growth Excretion Reproduction Deat/Life span
Hashim
What makes children from the same father and mother sometimes don't look alike?
Hashim
identification of problems
Nana Reply
what happens in the process of raising the human arms
Nana
what is biology
Brandi Reply
first step in scientific method
Brandi
In an investigation the pancreatic duct of a mammal was blocked.It was found that the blood sugar regulation remained normal while food digestion was impaired.Explain
Mac Reply
To begin with, obstruction of pancreatic duct will alter the blood sugar level as the juices responsible for glucose regulation will be rendered inconsequential. This will in turn affect the rate of digestion and absorbtion of digested food substances by the Villus .
Muktar
characteristics of algae
OMIME Reply
Algae are eukaryotic organisms. Algae do not have roots and stems. Algae have chlorophyll and helps in carrying out photosynthesis.
Aditi
Cell wall is the rigid layer enclosed by membranes of plants and prokayortic cell, it maintains the shape of the cell and serve as a protective barrier.
chizoba Reply
ECOLOGY: is a branch of biology that studies the interactions among organisms and their biophysical environment, which includes both biotic and abiotic components. 
chizoba
via nutrient cycles and energy flows. For instance, the energy from the sun is captured by plants through photosynthesis. Photosynthesis is a biological process through which plants manufacture their own food with the aid of light from the sun and frc sources (e.g. cabon dioxide and water)
chizoba
What is cell wall
Taiwo Reply
cell wall is the outemost rigid covering of the plants ,that provides protection to the plants.
Aditi
what is ecology, ecosystem?
Nkeng Reply
what is digestive system
Lucky Reply
digestive system is the human syman system that icludes esopuges stomach o braking down of food in to useful substance to our body
samrawit
definition of biology basics
Ritu Reply
the potential energy of a molecule can be inquired by their number of?
Jesus Reply
what is the full meaning of RNA
Ayo Reply
ribose nucleic acid
Nikita
Ribonucleic acid
Jesus
Ribo Nucleic Acid
Aditi
ribonucleic acid
Nana
discuss, describe at least three (3) methods that could be used to improve photosynthesis..
Marvel Reply
Improve the efficiency with which plants capture light Improve the efficiency by which plants turn light into energy The smart canopy concept develop crop planting schemes that increase the penetration of sunlight into lower-level leaves.
Jesus
what is osmosis
Aon Reply
movement of water molecule from higher to lower concentration through a semipereable membrene.
Dr
what of in the case of solute
Aon
osmosis is the movement of molecules from higher concentration region to lower concentration region through semi-permeable membrane.
Broad
in case of solute means that water moves from the region with lower solutes to the region with higher solute. so it is vice versa to water.
Broad
osmosis is the movement of water molecule from a region of lower concentration to a region of higher concentration through a semi permeable membrane
Nana

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Source:  OpenStax, Concepts of biology. OpenStax CNX. Feb 29, 2016 Download for free at http://cnx.org/content/col11487/1.9
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