Ligases
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DNA ligases mediate the sealing of single stranded breaks or gaps in DNA molecules by catalyzing the formation of phosphodiester bonds between DNA fragments, acting as molecular glue to join DNA fragments together. There are two main types of DNA ligases, E. coli ligase which requires NAD as a cofactor and T4 DNA ligase which uses ATP as a cofactor.
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Types of Restriction Endonuclease enzymes
Restriction enzymes are produced by bacterial cells to protect themselves from foreign DNA. There are three main activities of restriction enzymes: recognition, digestion, and modification. Restriction enzymes are classified into three groups based on their source and biochemical activity: isoschizomers which have the same recognition and cleavage sites, neoschizomers which have the same recognition site but different cleavage sites, and isocaudomers which have different recognition sites but the same cleavage sites.
Methanotrophs
Methanotrophs are prokaryotes that metabolize methane as their sole carbon and energy source. They can be bacteria or archaea and grow aerobically or anaerobically. Methanotrophs are common in environments where methane is produced, such as wetlands, soil, marshes, and landfills. They play an important role in oxidizing methane in the natural world and reducing methane emissions. Methanotrophs contain methane monooxygenase enzymes which allow them to utilize methane and also degrade various organic pollutants, making them useful for bioremediation applications.
Molecular biology, principle of immunization by L.pasteur and E.jenner.
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This document discusses reverse transcriptase and restriction enzymes. Reverse transcriptase is an enzyme that synthesizes DNA from an mRNA template. It is found in retroviruses and is used to make cDNA from mRNA. Restriction enzymes are molecular scissors found in bacteria that recognize and cut DNA at specific nucleotide sequences. Over 3000 restriction enzymes have been identified that can cut DNA into fragments for uses like DNA sequencing, cloning, and transformation. DNA ligase is then used to join DNA fragments back together.
Ligase enzyme
DNA ligase is an enzyme that catalyzes the formation of phosphodiester bonds between DNA fragments, joining two DNA strands together. It plays an important role in DNA replication by joining Okazaki fragments and filling in gaps, as well as in DNA repair and genetic engineering techniques like cloning. The most commonly used DNA ligase is from bacteriophage T4, which utilizes ATP as a cofactor and works efficiently at lower temperatures to ligate DNA strands with either sticky or blunt ends.
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This document discusses the history and properties of plasmids. Plasmids are extrachromosomal DNA molecules that are able to replicate independently of a cell's chromosomal DNA. They were first observed in bacteria in the early 1950s and play important roles in processes like antibiotic resistance and virulence. The document outlines the early studies that helped characterize plasmids and describes some of their key properties, such as their circular structure and ability to exist in different conformations. It also discusses how plasmids are used as cloning vectors to amplify genes and produce proteins for applications in research, medicine, and agriculture.
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Ligase enzyme
DNA ligase is an enzyme that catalyzes the formation of phosphodiester bonds between DNA fragments, joining two DNA strands together. It plays an important role in DNA replication by joining Okazaki fragments and filling in gaps, as well as in DNA repair and genetic engineering techniques like cloning. The most commonly used DNA ligase is from bacteriophage T4, which utilizes ATP as a cofactor and works efficiently at lower temperatures to ligate DNA strands with either sticky or blunt ends.
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A class of enzymes that alter the supercoiling of double-stranded DNA. (In supercoiling the DNA molecule coils up like a telephone cord, which shortens the molecule.) The topoisomerases act by transiently cutting one or both strands of the DNA.
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Since we are talking about DNA Methylases these are those enzymes in which the target or recipient molecule happens to be a DNA or Nucleotide in a DNA sequence.
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Archive contain atomic coordinates, bibliographic citations, primary and secondary structure information, crystallographic structure factors, NMR experimental data.
Hammerhead ribozyme
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SOS response was discovered by Miroslav Radman. It's a part of DNA repair system- synthesizes enzymes required for DNA repair. Cellular response to UV damage.
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Methanotrophs are unique bacteria that use methane as their sole carbon and energy source. They play several important roles in the environment. First, they oxidize methane produced by archaea, reducing the amount of this potent greenhouse gas that enters the atmosphere. Second, they can degrade various organic pollutants using monooxygenase enzymes. Finally, they can remediate heavy metals by transforming more toxic forms into less toxic forms or facilitating precipitation. Methanotrophs thus aid in carbon and pollutant cycling while also helping to mitigate climate change.
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Stirred Tank Bioreactor: Versatile and widely used for efficient mixing and oxygen transfer in microbial cultures and large-scale production.
Airlift Bioreactor: Ideal for gentle mixing in shear-sensitive and mammalian cell cultures using gas injection.
Fluidized Bed Bioreactor: Offers a large surface area for cell attachment, suitable for immobilized cell cultures or biofilm formation.
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Membrane Bioreactor: Combines biological treatment with membrane filtration for effective contaminant removal in wastewater treatment applications.
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This document discusses the application of biotechnology in agriculture, focusing on genetically modified crops. It provides examples of GM crops including Bt cotton, Bt brinjal, and golden rice. Bt crops are engineered to produce insecticidal Cry proteins from Bacillus thuringiensis to resist pests. Golden rice is genetically modified to produce beta-carotene and address vitamin A deficiency. The document also discusses the Flavr Savr tomato, the first commercially available GM food, which was engineered for longer shelf life by inhibiting the polygalacturonase enzyme. Finally, it introduces the concept of edible vaccines, where plants are engineered to produce vaccine proteins that are consumed orally.
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Application of biotechnology in medicine
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Methods of gene transfer
Gene transfer techniques can be direct or indirect. Direct techniques introduce foreign DNA into plant cells without a biological agent, using methods like microinjection, microprojectiles, protoplast fusion, electroporation, and polyethylene glycol treatment. Indirect gene transfer uses the bacterium Agrobacterium tumefaciens, which transfers DNA (T-DNA) from its tumor-inducing plasmid into the host plant genome, allowing genetic modification of plants. Techniques like bacterial transformation and transduction can also directly transfer genes between bacteria using viruses or naked DNA. Overall, a variety of methods have been developed to introduce foreign genes into organisms and achieve genetic modification.
Southern blotting
Southern blotting is a technique used to detect specific DNA sequences. It involves separating DNA fragments by size via gel electrophoresis, transferring them to a membrane, and using a labeled probe to hybridize to the target sequence. The probe binds only to complementary DNA fragments, allowing their detection and location on the membrane via autoradiography. Key steps include restriction enzyme digestion of DNA, gel electrophoresis, membrane transfer, probe hybridization, and x-ray film development to visualize hybridized fragments. Southern blotting is used for applications like mutation detection, gene rearrangement studies, and forensic analysis.
Restriction endonuclease
Restriction endonucleases are enzymes produced by bacteria that recognize specific sequences in foreign DNA and cut the DNA at or near the recognition sites. They function as part of the bacterial immune system to degrade foreign DNA. Restriction endonucleases cut DNA into fragments that have either blunt or sticky ends, depending on where they cut the DNA strands. They are named based on the bacteria that produces them and are classified into three types depending on their cleavage patterns. Type II restriction endonucleases cut directly at their recognition sites and produce fragments with sticky or blunt ends, making them useful for genetic engineering applications.
Polymerase chain reaction (pcr)
PCR is a technique used to amplify a specific segment of DNA. It involves repeated cycles of heating and cooling of the DNA sample in the presence of DNA polymerase, primers, and nucleotides. During each cycle, the DNA denatures, the primers anneal to the DNA, and the polymerase extends the primers to make copies of the DNA segment. This allows millions of copies of the target DNA sequence to be generated from a very small initial sample. PCR is widely used in applications like disease diagnosis, DNA fingerprinting, and molecular cloning.
Gene cloning
Gene cloning involves producing multiple copies of a gene using genetic engineering techniques. The main steps are: isolating the donor DNA fragment, inserting it into a vector, transforming host cells, and selecting recombinant cells containing the desired DNA insert. Gene cloning strategies include creating genomic DNA libraries, which contain all DNA from an organism's genome, and cDNA libraries, which contain only coding regions created from mRNA. Genomic libraries contain both coding and non-coding regions from genomes, while cDNA libraries contain only coding exons synthesized from mRNA using reverse transcriptase.
rDna technology
rDNA technology involves deliberately manipulating the genetic makeup of cells or organisms in a controlled way. Its major goals are to form gene libraries, isolate and amplify genes of interest, and alter genetic makeup and gene expression patterns. The key steps are isolating DNA from cells, fragmenting the DNA with restriction enzymes, amplifying the gene of interest with PCR, and inserting the recombinant DNA into a host with enzymes.
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