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EHA101
EHA101 was one of the first and most widely used ''Agrobacterium'' helper plasmid for plant gene transfer. Created in 1985 in the laboratory of Mary-Dell Chilton at Washington University in St. Louis, it was named after the graduate student who constructed it. The EH stands for "Elizabeth Hood" and A for "''Agrobacterium''". The EHA101 helper strain is a derivative of A281, the hypervirulent ''A. tumefaciens'' strain that causes large, fast-growing tumors on solanaceous plants. This strain is used for moving genes of interest into many hundreds of species of plants all over the world. For recalcitrant crops such as maize, wheat, and rice, the EHA helper strains are often employed for gene transfer. These strains are efficient at promoting T-DNA transfer because of the hypervirulence of the vir genes suggesting that a higher success rate can be achieved on these "hard to transform" crops or cultivars. The chromosomal background of EHA101 is C58C1, a cured nopaline strain. The h ...
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T-DNA Binary System
A transfer DNA (T-DNA) binary system is a pair of plasmids consisting of a T-DNA binary vector and a ''vir'' helper plasmid. The two plasmids are used together (thus ''binary'') to produce genetically modified plants. They are artificial vectors that have been derived from the naturally occurring Ti plasmid found in bacterial species of the genus ''Agrobacterium'', such as '' A. tumefaciens''. The binary vector is a ''shuttle vector'', so-called because it is able to replicate in multiple hosts (e.g. '' Escherichia coli'' and ''Agrobacterium''). Systems in which T-DNA and ''vir'' genes are located on separate replicons are called T-DNA binary systems. T-DNA is located on the binary vector (the non-T-DNA region of this vector containing origin(s) of replication that could function both in ''E. coli'' and ''Agrobacterium'', and antibiotic resistance genes used to select for the presence of the binary vector in bacteria, became known as vector backbone sequences). The replicon conta ...
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Agrobacterium
''Agrobacterium'' is a genus of Gram-negative bacteria established by H. J. Conn that uses horizontal gene transfer to cause tumors in plants. ''Agrobacterium tumefaciens'' is the most commonly studied species in this genus. ''Agrobacterium'' is well known for its ability to transfer DNA between itself and plants, and for this reason it has become an important tool for genetic engineering. Nomenclatural History Leading up to the 1990s, the genus ''Agrobacterium'' was used as a wastebasket taxon. With the advent of 16S sequencing, many ''Agrobacterium'' species (especially the marine species) were reassigned to genera such as ''Ahrensia'', ''Pseudorhodobacter'', ''Ruegeria'', and ''Stappia''. The remaining ''Agrobacterium'' species were assigned to three biovars: biovar 1 (''Agrobacterium tumefaciens''), biovar 2 (''Agrobacterium rhizogenes''), and biovar 3 (''Agrobacterium vitis''). In the early 2000s, ''Agrobacterium'' was synonymized with the genus ''Rhizobium''. This move pr ...
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Site-directed Mutagenesis
Site-directed mutagenesis is a molecular biology method that is used to make specific and intentional mutating changes to the DNA sequence of a gene and any gene products. Also called site-specific mutagenesis or oligonucleotide-directed mutagenesis, it is used for investigating the structure and biological activity of DNA, RNA, and protein molecules, and for protein engineering. Site-directed mutagenesis is one of the most important laboratory techniques for creating DNA libraries by introducing mutations into DNA sequences. There are numerous methods for achieving site-directed mutagenesis, but with decreasing costs of oligonucleotide synthesis, artificial gene synthesis is now occasionally used as an alternative to site-directed mutagenesis. Since 2013, the development of the CRISPR/Cas9 technology, based on a prokaryotic viral defense system, has also allowed for the editing of the genome, and mutagenesis may be performed ''in vivo'' with relative ease. History Early attempt ...
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Antibiotic Resistance
Antimicrobial resistance (AMR) occurs when microbes evolve mechanisms that protect them from the effects of antimicrobials. All classes of microbes can evolve resistance. Fungi evolve antifungal resistance. Viruses evolve antiviral resistance. Protozoa evolve antiprotozoal resistance, and bacteria evolve antibiotic resistance. Those bacteria that are considered extensively drug resistant (XDR) or totally drug-resistant (TDR) are sometimes called "superbugs".A.-P. Magiorakos, A. Srinivasan, R. B. Carey, Y. Carmeli, M. E. Falagas, C. G. Giske, S. Harbarth, J. F. Hinndler ''et al''Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria... Clinical Microbiology and Infection, Vol 8, Iss. 3 first published 27 July 2011 ia Wiley Online Library Retrieved 28 August 2020 Although antimicrobial resistance is a naturally-occurring process, it is often the result of improper usage of the drugs and management of the infections. Antibiotic resistance is a major subset ...
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Kanamycin
Kanamycin A, often referred to simply as kanamycin, is an antibiotic used to treat severe bacterial infections and tuberculosis. It is not a first line treatment. It is used by mouth, injection into a vein, or injection into a muscle. Kanamycin is recommended for short-term use only, usually from 7 to 10 days. As with most antibiotics, it is ineffective in viral infections. Common side effects include hearing and balance problems. Kidney problems may also occur. Kanamycin is not recommended during pregnancy as it may harm the baby. It is likely safe during breastfeeding. Kanamycin is in the aminoglycoside family of medications. It works by blocking the production of proteins that are required for bacterial survival. Kanamycin was first isolated in 1957 by Hamao Umezawa from the bacterium ''Streptomyces kanamyceticus''. It was removed from the World Health Organization's List of Essential Medicines in 2019. It is no longer marketed in the United States. Medical uses Spectrum ...
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Nopaline
Nopaline is a chemical compound derived from the amino acids glutamic acid and arginine. It is classified as an opine. Ti plasmids are classified on the basis of the different types of opines they produce. These may be nopaline plasmids, octopine plasmids and agropine plasmids. These opines are condensation products of amino acids and keto acids or may be derived from sugars. The opines are used as carbon and nitrogen sources and metabolized by ''Agrobacterium ''Agrobacterium'' is a genus of Gram-negative bacteria established by H. J. Conn that uses horizontal gene transfer to cause tumors in plants. '' Agrobacterium tumefaciens'' is the most commonly studied species in this genus. ''Agrobacterium'' i ...''. References Amino acid derivatives {{organic-compound-stub ...
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Cultivars
A cultivar is a type of Horticulture, cultivated plant that people have selected for desired phenotypic trait, traits and when Plant propagation, propagated retain those traits. Methods used to propagate cultivars include: division, root and stem cuttings, offsets, grafting, micropropagation, tissue culture, or carefully controlled seed production. Most cultivars arise from purposeful human genetic engineering, manipulation, but some originate from wild plants that have distinctive characteristics. Cultivar names are chosen according to rules of the International Code of Nomenclature for Cultivated Plants (ICNCP), and not all cultivated plants qualify as cultivars. Horticulturists generally believe the word ''cultivar''''Cultivar'' () has two meanings, as explained in ''#Formal definition, Formal definition'': it is a classification category and a taxonomic unit within the category. When referring to a taxon, the word does not apply to an individual plant but to all plants that s ...
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Ti Plasmid
A tumour inducing (Ti) plasmid is a plasmid found in pathogenic species of ''Agrobacterium'', including ''A. tumefaciens, ''A. rhizogenes'', ''A. rubi'' and ''A. vitis''. Evolutionarily, the Ti plasmid is part of a family of plasmids carried by many species of Alphaproteobacteria. Members of this plasmid family are defined by the presence of a conserved DNA region known as the ''repABC'' gene cassette, which mediates the replication of the plasmid, the partitioning of the plasmid into daughter cells during cell division as well as the maintenance of the plasmid at low copy numbers in a cell. The Ti plasmids themselves are sorted into different categories based on the type of molecule, or opine, they allow the bacteria to break down as an energy source. The presence of this Ti plasmid is essential for the bacteria to cause crown gall disease in plants. This is facilitated via certain crucial regions in the Ti plasmid, including the ''vir'' region, which encodes for virulence g ...
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Wheat
Wheat is a grass widely cultivated for its seed, a cereal grain that is a worldwide staple food. The many species of wheat together make up the genus ''Triticum'' ; the most widely grown is common wheat (''T. aestivum''). The archaeological record suggests that wheat was first cultivated in the regions of the Fertile Crescent around 9600 BCE. Botanically, the wheat kernel is a type of fruit called a caryopsis. Wheat is grown on more land area than any other food crop (, 2014). World trade in wheat is greater than for all other crops combined. In 2020, world production of wheat was , making it the second most-produced cereal after maize. Since 1960, world production of wheat and other grain crops has tripled and is expected to grow further through the middle of the 21st century. Global demand for wheat is increasing due to the unique viscoelastic and adhesive properties of gluten proteins, which facilitate the production of processed foods, whose consumption is inc ...
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Oryza Sativa
''Oryza sativa'', commonly known as Asian rice or indica rice, is the plant species most commonly referred to in English as ''rice''. It is the type of farmed rice whose cultivars are most common globally, and was first domesticated in the Yangtze River basin in China 13,500 to 8,200 years ago. ''Oryza sativa'' belongs to the genus '' Oryza'' of the grass family Poaceae. With a genome consisting of 430 Mbp across 12 chromosomes, it is renowned for being easy to genetically modify and is a model organism for the botany of cereals. Classification ''Oryza sativa'' contains two major subspecies: the sticky, short-grained ''japonica'' or ''sinica'' variety, and the nonsticky, long-grained ' rice variety. ''Japonica'' was domesticated in the Yangtze Valley 9–6,000 years ago, and its varieties can be cultivated in dry fields (it is cultivated mainly submerged in Japan), in temperate East Asia, upland areas of Southeast Asia, and high elevations in South Asia, while ''indica'' w ...
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Maize
Maize ( ; ''Zea mays'' subsp. ''mays'', from es, maíz after tnq, mahiz), also known as corn (North American and Australian English), is a cereal grain first domesticated by indigenous peoples in southern Mexico about 10,000 years ago. The leafy stalk of the plant produces pollen inflorescences (or "tassels") and separate ovuliferous inflorescences called ears that when fertilized yield kernels or seeds, which are fruits. The term ''maize'' is preferred in formal, scientific, and international usage as a common name because it refers specifically to this one grain, unlike ''corn'', which has a complex variety of meanings that vary by context and geographic region. Maize has become a staple food in many parts of the world, with the total production of maize surpassing that of wheat or rice. In addition to being consumed directly by humans (often in the form of masa), maize is also used for corn ethanol, animal feed and other maize products, such as corn starch and ...
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Recalcitrant
Recalcitrant seeds are seeds that do not survive drying and freezing during ex-situ conservation. By and large, these seeds cannot resist the effects of drying or temperatures less than 10 °C (50 °F); thus, they cannot be stored for long periods like orthodox seeds because they can lose their viability. Plants that produce recalcitrant seeds include avocado, mango, mangosteen, lychee, cocoa, rubber tree, some horticultural trees, aquatic plants such as ''Nymphaea caerulea'''','' and several plants used in traditional medicine, such as species of ''Virola'' and ''Pentaclethra''. Generally speaking, most tropical pioneer species have orthodox seeds but many climax species have recalcitrant or intermediate seeds. Mechanisms of damage The two main mechanisms causing damage to recalcitrant seeds are desiccation effects on the intracellular structures and metabolic damage from the formation of toxic chemicals such as free radicals. An example of the first type of damage would be ...
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