- published: 25 Nov 2013
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Deoxyribonucleic acid (i/diˈɒksiˌraɪboʊnjʊˌkliːɪk, -ˌkleɪɪk/;DNA) is a molecule that carries most of the genetic instructions used in the development, functioning and reproduction of all known living organisms and many viruses. DNA is a nucleic acid; alongside proteins and carbohydrates, nucleic acids compose the three major macromolecules essential for all known forms of life. Most DNA molecules consist of two biopolymer strands coiled around each other to form a double helix. The two DNA strands are known as polynucleotides since they are composed of simpler units called nucleotides. Each nucleotide is composed of a nitrogen-containing nucleobase—either cytosine (C), guanine (G), adenine (A), or thymine (T)—as well as a monosaccharide sugar called deoxyribose and a phosphate group. The nucleotides are joined to one another in a chain by covalent bonds between the sugar of one nucleotide and the phosphate of the next, resulting in an alternating sugar-phosphate backbone. According to base pairing rules (A with T, and C with G), hydrogen bonds bind the nitrogenous bases of the two separate polynucleotide strands to make double-stranded DNA. The total amount of related DNA base pairs on Earth is estimated at 5.0 x 1037, and weighs 50 billion tonnes. In comparison, the total mass of the biosphere has been estimated to be as much as 4 TtC (trillion tons of carbon).
In molecular biology, DNA ligase is a specific type of enzyme, a ligase, (EC 6.5.1.1) that facilitates the joining of DNA strands together by catalyzing the formation of a phosphodiester bond. It plays a role in repairing single-strand breaks in duplex DNA in living organisms, but some forms (such as DNA ligase IV) may specifically repair double-strand breaks (i.e. a break in both complementary strands of DNA). Single-strand breaks are repaired by DNA ligase using the complementary strand of the double helix as a template, with DNA ligase creating the final phosphodiester bond to fully repair the DNA.
DNA ligase is used in both DNA repair and DNA replication (see Mammalian ligases). In addition, DNA ligase has extensive use in molecular biology laboratories for recombinant DNA experiments (see Applications in molecular biology research). Purified DNA ligase is used in gene cloning to join DNA molecules together to form recombinant DNA.
The mechanism of DNA ligase is to form two covalent phosphodiester bonds between 3' hydroxyl ends of one nucleotide, ("acceptor") with the 5' phosphate end of another ("donor"). ATP is required for the ligase reaction, which proceeds in three steps:
Khan Academy is a non-profit educational organization created in 2006 by educator Salman Khan with the aim of providing a free, world-class education for anyone, anywhere. The organization produces short lectures in the form of YouTube videos. In addition to micro lectures, the organization's website features practice exercises and tools for educators. All resources are available for free to anyone around the world. The main language of the website is English, but the content is also available in other languages.
The founder of the organization, Salman Khan, was born in New Orleans, Louisiana, United States to immigrant parents from Bangladesh and India. After earning three degrees from the Massachusetts Institute of Technology (a BS in mathematics, a BS in electrical engineering and computer science, and an MEng in electrical engineering and computer science), he pursued an MBA from Harvard Business School.
In late 2004, Khan began tutoring his cousin Nadia who needed help with math using Yahoo!'s Doodle notepad.When other relatives and friends sought similar help, he decided that it would be more practical to distribute the tutorials on YouTube. The videos' popularity and the testimonials of appreciative students prompted Khan to quit his job in finance as a hedge fund analyst at Connective Capital Management in 2009, and focus on the tutorials (then released under the moniker "Khan Academy") full-time.
The DNA polymerases are enzymes that create DNA molecules by assembling nucleotides, the building blocks of DNA. These enzymes are essential to DNA replication and usually work in pairs to create two identical DNA strands from a single original DNA molecule. During this process, DNA polymerase “reads” the existing DNA strands to create two new strands that match the existing ones.
Every time a cell divides, DNA polymerase is required to help duplicate the cell’s DNA, so that a copy of the original DNA molecule can be passed to each of the daughter cells. In this way, genetic information is transmitted from generation to generation.
Before replication can take place, an enzyme called helicase unwinds the DNA molecule from its tightly woven form. This opens up or “unzips” the double-stranded DNA to give two single strands of DNA that can be used as templates for replication.
In 1956, Arthur Kornberg and colleagues discovered the enzyme DNA polymerase I, also known as Pol I, in Escherichia coli. They described the DNA replication process by which DNA polymerase copies the base sequence of a template DNA strand. Subsequently, in 1959, Kornberg was awarded the Nobel Prize in Physiology or Medicine for this work.DNA polymerase II was also discovered by Kornberg and Malcolm E. Gefter in 1970 while further elucidating the role of Pol I in E. coli DNA replication.
DNA replication is the process of producing two identical replicas from one original DNA molecule. This biological process occurs in all living organisms and is the basis for biological inheritance. DNA is made up of two strands and each strand of the original DNA molecule serves as a template for the production of the complementary strand, a process referred to as semiconservative replication. Cellular proofreading and error-checking mechanisms ensure near perfect fidelity for DNA replication.
In a cell, DNA replication begins at specific locations, or origins of replication, in the genome. Unwinding of DNA at the origin and synthesis of new strands results in replication forks growing bidirectional from the origin. A number of proteins are associated with the replication fork which helps in terms of the initiation and continuation of DNA synthesis. Most prominently, DNA polymerase synthesizes the new DNA by adding complementary nucleotides to the template strand.
DNA replication can also be performed in vitro (artificially, outside a cell). DNA polymerases isolated from cells and artificial DNA primers can be used to initiate DNA synthesis at known sequences in a template DNA molecule. The polymerase chain reaction (PCR), a common laboratory technique, cyclically applies such artificial synthesis to amplify a specific target DNA fragment from a pool of DNA.
This lecture about DNA ligase explains how does DNA ligase works. http://shomusbiology.com/ Download the study materials here- http://shomusbiology.weebly.com/bio-materials.html In molecular biology, DNA ligase is a specific type of enzyme, a ligase, (EC 6.5.1.1) that facilitates the joining of DNA strands together by catalyzing the formation of a phosphodiester bond. It plays a role in repairing single-strand breaks in duplex DNA in living organisms, but some forms (such as DNA ligase IV) may specifically repair double-strand breaks (i.e. a break in both complementary strands of DNA). Single-strand breaks are repaired by DNA ligase using the complementary strand of the double helix as a template,[1] with DNA ligase creating the final phosphodiester bond to fully repair the DNA. DNA ligas...
Ligation, the process of joining DNA fragments with a DNA ligase, proceeds in three steps. Learn more about DNA ligation at https://www.neb.com/applications/dna-modification/dna-ligation
This lecture explains the DNA ligase mechanism of t4 DNA ligase. http://shomusbiology.weebly.com/ Download the study materials here- http://shomusbiology.weebly.com/bio-materials.html Molecular cloning is the laboratory process used to create recombinant DNA.[1][2][3][4] It is one of two widely used methods (along with polymerase chain reaction, abbr. PCR) used to direct the replication of any specific DNA sequence chosen by the experimentalist. The fundamental difference between the two methods is that molecular cloning involves replication of the DNA within a living cell, while PCR replicates DNA in the test tube, free of living cells. Formation of recombinant DNA requires a cloning vector, a DNA molecule that will replicate within a living cell. Vectors are generally derived from plasm...
Detailed review of DNA ligase and joining of Okazaki fragments on the lagging strand
3.4.1 Explain DNA Replication DNA Replication is the process by which two identical pieces of DNA are produced from one original piece of DNA. This process ensures that new cells that are made contain exactly the same information as the original cell. Firstly you need to recall the structure of DNA in order to understand the process of replication. Recall that the molecule is double stranded and that those strand are antiparallel to each other. You can see this on the video by the labeled prime ends. Please refer to the video regarding the structure of DNA (3.3.3/3.3.4/3.3.5) in order to familiarize yourself with its structure if you need to refresh! The two strands undergo replication in a slightly different fashion so in the video I show you first the leading strand only followed ...
This is an audioblog of the article "How DNA ligase works" from Bitesize Bio
▶ This video channel is developed by Amrita University's CREATE http://www.amrita.edu/create ▶ Subscribe @ https://www.youtube.com/user/amritacreate http://www.youtube.com/amritavlab ▶ Like us @ https://www.facebook.com/CREATEatAmrita ▶ For more Information @ http://vlab.amrita.edu/index.php?sub=3&brch;=186&sim=781&cnt;=1 ▶ Amrita Virtual Lab Project website http://vlab.amrita.edu DNA ligation is the act of joining together DNA strands with covalent bonds with the aim of making new viable DNA or plasmids. The enzyme used to ligate DNA fragments is T4 DNA ligase, which originates from the T4 bacteriophage. This enzyme will ligate DNA fragments having overhanging, cohesive ends that are annealed together. A ligation reaction requires three ingredients in addition to water: 1. Tw...
DNA Ligase or 'molecular gums' or 'joining enzymes': Definition and function in rDNA technology
Introduction to DNA cloning. Watch the next lesson: https://www.khanacademy.org/test-prep/mcat/biomolecules/dna-technology/v/hybridization-microarray?utm_source=YT&utm;_medium=Desc&utm;_campaign=mcat Missed the previous lesson? https://www.khanacademy.org/test-prep/mcat/biomolecules/dna-technology/v/dna-libraries-generating-cdna?utm_source=YT&utm;_medium=Desc&utm;_campaign=mcat MCAT on Khan Academy: Go ahead and practice some passage-based questions! About Khan Academy: Khan Academy offers practice exercises, instructional videos, and a personalized learning dashboard that empower learners to study at their own pace in and outside of the classroom. We tackle math, science, computer programming, history, art history, economics, and more. Our math missions guide learners from kindergarten to...
This lecture about DNA ligase explains how does DNA ligase works. http://shomusbiology.com/ Download the study materials here- http://shomusbiology.weebly.com/bio-materials.html In molecular biology, DNA ligase is a specific type of enzyme, a ligase, (EC 6.5.1.1) that facilitates the joining of DNA strands together by catalyzing the formation of a phosphodiester bond. It plays a role in repairing single-strand breaks in duplex DNA in living organisms, but some forms (such as DNA ligase IV) may specifically repair double-strand breaks (i.e. a break in both complementary strands of DNA). Single-strand breaks are repaired by DNA ligase using the complementary strand of the double helix as a template,[1] with DNA ligase creating the final phosphodiester bond to fully repair the DNA. DNA ligas...
Ligation, the process of joining DNA fragments with a DNA ligase, proceeds in three steps. Learn more about DNA ligation at https://www.neb.com/applications/dna-modification/dna-ligation
This lecture explains the DNA ligase mechanism of t4 DNA ligase. http://shomusbiology.weebly.com/ Download the study materials here- http://shomusbiology.weebly.com/bio-materials.html Molecular cloning is the laboratory process used to create recombinant DNA.[1][2][3][4] It is one of two widely used methods (along with polymerase chain reaction, abbr. PCR) used to direct the replication of any specific DNA sequence chosen by the experimentalist. The fundamental difference between the two methods is that molecular cloning involves replication of the DNA within a living cell, while PCR replicates DNA in the test tube, free of living cells. Formation of recombinant DNA requires a cloning vector, a DNA molecule that will replicate within a living cell. Vectors are generally derived from plasm...
Detailed review of DNA ligase and joining of Okazaki fragments on the lagging strand
3.4.1 Explain DNA Replication DNA Replication is the process by which two identical pieces of DNA are produced from one original piece of DNA. This process ensures that new cells that are made contain exactly the same information as the original cell. Firstly you need to recall the structure of DNA in order to understand the process of replication. Recall that the molecule is double stranded and that those strand are antiparallel to each other. You can see this on the video by the labeled prime ends. Please refer to the video regarding the structure of DNA (3.3.3/3.3.4/3.3.5) in order to familiarize yourself with its structure if you need to refresh! The two strands undergo replication in a slightly different fashion so in the video I show you first the leading strand only followed ...
This is an audioblog of the article "How DNA ligase works" from Bitesize Bio
▶ This video channel is developed by Amrita University's CREATE http://www.amrita.edu/create ▶ Subscribe @ https://www.youtube.com/user/amritacreate http://www.youtube.com/amritavlab ▶ Like us @ https://www.facebook.com/CREATEatAmrita ▶ For more Information @ http://vlab.amrita.edu/index.php?sub=3&brch;=186&sim=781&cnt;=1 ▶ Amrita Virtual Lab Project website http://vlab.amrita.edu DNA ligation is the act of joining together DNA strands with covalent bonds with the aim of making new viable DNA or plasmids. The enzyme used to ligate DNA fragments is T4 DNA ligase, which originates from the T4 bacteriophage. This enzyme will ligate DNA fragments having overhanging, cohesive ends that are annealed together. A ligation reaction requires three ingredients in addition to water: 1. Tw...
DNA Ligase or 'molecular gums' or 'joining enzymes': Definition and function in rDNA technology
Introduction to DNA cloning. Watch the next lesson: https://www.khanacademy.org/test-prep/mcat/biomolecules/dna-technology/v/hybridization-microarray?utm_source=YT&utm;_medium=Desc&utm;_campaign=mcat Missed the previous lesson? https://www.khanacademy.org/test-prep/mcat/biomolecules/dna-technology/v/dna-libraries-generating-cdna?utm_source=YT&utm;_medium=Desc&utm;_campaign=mcat MCAT on Khan Academy: Go ahead and practice some passage-based questions! About Khan Academy: Khan Academy offers practice exercises, instructional videos, and a personalized learning dashboard that empower learners to study at their own pace in and outside of the classroom. We tackle math, science, computer programming, history, art history, economics, and more. Our math missions guide learners from kindergarten to...
In this webinar, NEB Scientist and ligase expert Greg Lohman discusses mismatch ligation by DNA ligases and the molecular diagnostics applications that depend on the use of high-fidelity DNA ligases like NEB’s HiFi Taq DNA Ligase to detect single base differences in DNA. Learn more about HiFi Ligase at www.neb.com/M0647
In this video I have explained DNA replication in prokaryotes in detail. I have explained about all the enzymes and proteins involved such as DnaA protein, helicases, SSB, primase, DNA pol III, DNA pol I, DNA ligase, topoisomerase II and IV etc. At 4:48, I have explained the OriC or origin of replication. At 17:57, I have explained leading and lagging strand. At 28:27, I have listed all the proteins and enzymes involved in replication in order. SUBSCRIBE TO THE CHANNEL FOR NEW VIDEOS EVERY WEEK :) http://www.youtube.com/c/NowIKnow13 Facebook: https://www.facebook.com/nowiknow13 Twitter: https://twitter.com/rupalgogia Now I Know Blog: http://rupalgogia.blogspot.in/
This lecture explains about the cDNA library construction and screening. http://shomusbiology.weebly.com/ Download the study materials here- http://shomusbiology.weebly.com/bio-materials.html A cDNA library is a combination of cloned cDNA (complementary DNA) fragments inserted into a collection of host cells, which together constitute some portion of the transcriptome of the organism. cDNA is produced from fully transcribed mRNA found in the nucleus and therefore contains only the expressed genes of an organism. Similarly, tissue specific cDNA libraries can be produced. In eukaryotic cells the mature mRNA is already spliced, hence the cDNA produced lacks introns and can be readily expressed in a bacterial cell. While information in cDNA libraries is a powerful and useful tool since gene p...
Contact me at kgahern@davincipress.com Friend me on Facebook at kevin.g.ahern Highlights DNA Replication/Repair/Recombination IV 1. The linear ends of the chromosomes are called telomeres. Telomeric sequences have thousands of copies of repeats of short sequences. 2. The enzyme that builds telomeres is called telomerase and is found predominantly in fetal and cancer cells, as well as fertilized eggs. Differentiated cells for the most part do not appear to have an active telomerase. 3. With each round of DNA replication, linear chromosomes in eukaryotes shorten. Thus, the more telomeric sequences a chromosome has, the more divisions it can undergo before the telomeres are "eaten up". 4. Telomerase is a reverse transcriptase - an enzyme that uses an RNA template (a circular RNA that it ...
Contact me at kgahern@davincipress.com Facebook friend me at https://www.facebook.com/kevin.g.ahern Highlights Translation 3 1. Translation is a target for action of antibiotics. This works because the phenomenon of translation in prokaryotes is different enough from that of eukaryotes that specific inhibitors of prokaryotic translation can be found that have no effect on eukaryotic translation. As a result, prokaryotes can be killed without any effect on eukaryotes taking said antibiotics. One translational inhibitor of prokaryotic translation is the antibiotic puromycin, which acts to prematurely terminate prokaryotic translation at the elongation phase. This happens because puromycin looks like a tRNA and fits into the A (or P) site of the ribosome. In the A site, it can be attached ...
Email Kevin at kgahern@davincipress.com Facebook friend me at kevin.g.ahern DNA Replication/Repair/Recombination III 1. DNA Polymerase I has a 3' to 5' exonuclease that removes RNA primers as the DNA polymerase fills in the space behind it as it moves along. 2. DNA Polymerase III is not very abundant in an E. coli cell, but it is the polymerase that makes most of the cellular DNA. 3. E. coli DNA replication occurs at 1000 base pairs per second. This requires a machine turning at 5000 to 6000 rpm. E. coli's helicase protein unwinds DNA at a rate of at least 5000 - 6000 rpm. The protein separates strands so as to make single strands accessible for replication. Unwinding of strands causes superhelical tension to increase ahead of the helicase. Topoisomerase II (gyrase) relieves the tensi...
http://www.ibiology.org/ibioseminars/toto-olivera-part-1.html Talk Overview: Although snails are not the first animals that come to mind when venoms are mentioned, there are, in fact, thousands of species of venomous predatory marine snails. The most intensively studied of these are the ~700 species of cone snails (Conus). Each snail species has ~100 different peptide neurotoxins present in its venom. Thus, conus venoms are a source of over 100,000 pharmacologically active peptides. In Part 1 of his lecture, Olivera tells us how he first became interested in studying conus venom. His lab found that conus venom peptides are organized into groups or “cabals” each of which act on a group of related molecular targets such as ion channels or receptors. This venom complexity gives snails an...
1. Contact me at kgahern@davincipress.com / Friend me on Facebook (kevin.g.ahern) 2. Download my free biochemistry book at http://biochem.science.oregonstate.edu/biochemistry-free-and-easy 3. Take my free iTunes U course at https://itunes.apple.com/us/course/biochemistry/id556410409 4. Check out my free book for pre-meds at http://biochem.science.oregonstate.edu/biochemistry-free-and-easy 5. Lecturio videos for medical students - https://www.lecturio.com/medical-courses/biochemistry.course 6. Course video channel at http://www.youtube.com/user/oharow/videos?view=1 7. Check out all of my free workshops at http://oregonstate.edu/dept/biochem/ahern/123.html 8. Check out my Metabolic Melodies at http://www.davincipress.com/ 9. My courses can be taken for credit (wherever you live) via OSU's e...
http://www.ibiology.org/ibioseminars/toto-olivera-part-2.html Talk Overview: Although snails are not the first animals that come to mind when venoms are mentioned, there are, in fact, thousands of species of venomous predatory marine snails. The most intensively studied of these are the ~700 species of cone snails (Conus). Each snail species has ~100 different peptide neurotoxins present in its venom. Thus, conus venoms are a source of over 100,000 pharmacologically active peptides. In Part 1 of his lecture, Olivera tells us how he first became interested in studying conus venom. His lab found that conus venom peptides are organized into groups or “cabals” each of which act on a group of related molecular targets such as ion channels or receptors. This venom complexity gives snails an...
http://www.ibiology.org/ibioseminars/toto-olivera-part-3.html Talk Overview: Although snails are not the first animals that come to mind when venoms are mentioned, there are, in fact, thousands of species of venomous predatory marine snails. The most intensively studied of these are the ~700 species of cone snails (Conus). Each snail species has ~100 different peptide neurotoxins present in its venom. Thus, conus venoms are a source of over 100,000 pharmacologically active peptides. In Part 1 of his lecture, Olivera tells us how he first became interested in studying conus venom. His lab found that conus venom peptides are organized into groups or “cabals” each of which act on a group of related molecular targets such as ion channels or receptors. This venom complexity gives snails an...