Recombinant Dna Technology Process — a labelled NEET Biology diagram with a definitions lexicon.
Recombinant DNA Technology Process Labelled parts: Recombinant DNA technology, Restriction digestion, Restriction enzyme, EcoRI, Plasmid vector, pBR322, AmpR, TetR, DNA ligase, Gene of interest, Ligation, Transformation, E. coli, Selection, Antibiotic plate, PCR cycle, Taq polymerase, Denaturation, Annealing, Extension, Primers. This technique involves combining genetic material from two or more different sources to create a novel DNA molecule. It is used to produce genetically modified organisms (GMOs) for various applications like drug production. FYI: The process typically involves cutting DNA using restriction enzymes and joining it using DNA ligase. These are bacterial enzymes that recognize and cut the DNA backbone at specific, short sequences of nucleotides (recognition sites). They are crucial tools in molecular biology for gene cloning. FYI: Restriction enzymes often cut DNA in a staggered manner, leaving 'sticky ends' which facilitate joining with other DNA fragments. A plasmid is a small, circular, self-replicating piece of DNA found naturally in bacteria. A plasmid vector is a modified plasmid used in cloning to carry and replicate foreign DNA into a host cell. FYI: Vectors must contain an origin of replication (ori) to ensure they can be multiplied within the host cell. An enzyme that catalyzes the formation of a phosphodiester bond between two adjacent DNA fragments. It is essential for joining Okazaki fragments on the lagging strand during replication. FYI: DNA ligase requires ATP or NAD+ as an energy source to seal nicks in the DNA backbone. Ligation is the biochemical process of joining two or more DNA fragments together using the enzyme DNA ligase. This enzyme forms phosphodiester bonds between the sugar-phosphate backbones of the DNA strands. FYI: DNA ligase requires ATP (or NAD + ) as an energy source to catalyze the formation of the phosphodiester bond. This is the natural process by which a bacterial cell takes up naked exogenous DNA from its surrounding environment. In molecular biology, it is an artificial process used to introduce foreign DNA into competent bacterial cells. FYI: Competence is the physiological state of the bacterial cell that allows it to take up DNA, often induced by chemical treatments like CaCl 2 . Escherichia coli is a common bacterium found in the gut of warm-blooded animals. It is widely used in molecular biology as a model organism for genetic manipulation and protein expression. FYI: E. coli is often used as a host organism because it is fast-growing, easily cultured, and has a well-understood genetic machinery. An antibiotic plate is a culture medium (agar) containing a specific antibiotic substance. It is used to test the susceptibility of bacteria to various antibiotics. FYI: Bacteria that grow on the plate are susceptible, while those that do not grow (forming a clear zone) are resistant to the antibiotic. Polymerase Chain Reaction (PCR) is a technique used to amplify specific DNA sequences exponentially in vitro. A single cycle involves three steps: denaturation, annealing, and extension. FYI: The key enzyme used in PCR is Taq polymerase, which is thermostable and can withstand the high temperatures of denaturation.