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A DNA construct is an artificially-designed segment of DNA borne on a vector that can be used to incorporate genetic material into a target tissue or cell. [1] A DNA construct contains a DNA insert, called a transgene, delivered via a transformation vector which allows the insert sequence to be replicated and/or expressed in the target cell.
The vector itself generally carries a DNA sequence that consists of an insert (in this case the transgene) and a larger sequence that serves as the "backbone" of the vector. The purpose of a vector which transfers genetic information to another cell is typically to isolate, multiply, or express the insert in the target cell.
The two base-pair complementary chains of the DNA molecule allow replication of the genetic instructions. The "specific pairing" is a key feature of the Watson and Crick model of DNA, the pairing of nucleotide subunits. [5] In DNA, the amount of guanine is equal to cytosine and the amount of adenine is equal to thymine. The A:T and C:G pairs ...
The DNA is stored in a population of identical vectors, each containing a different insert of DNA. In order to construct a genomic library, the organism's DNA is extracted from cells and then digested with a restriction enzyme to cut the DNA into fragments of a specific size. The fragments are then inserted into the vector using DNA ligase. [1]
The Phusion DNA polymerase fills in any gaps where the fragments anneal. Taq DNA ligase repairs the nicks on both DNA strands. Because the T5 exonuclease is heat labile, it is inactivated at 50 °C after the initial chew back step. The product is thus stable, and the fragments assembled in the desired order.
The technical definition of a nucleosome includes a segment of DNA about 146 base pairs in length which makes 1.67 left-handed turns as it coils around the histone core, as well as a stretch of linker DNA (generally 38–80 bp) connecting it to an adjacent core particle, though the term is often used to refer to the core particle alone.
In the natural sciences, a vector quantity (also known as a vector physical quantity, physical vector, or simply vector) is a vector-valued physical quantity. [ 1 ] [ 2 ] It is typically formulated as the product of a unit of measurement and a vector numerical value ( unitless ), often a Euclidean vector with magnitude and direction .
An expression vector, otherwise known as an expression construct, is usually a plasmid or virus designed for gene expression in cells. The vector is used to introduce a specific gene into a target cell, and can commandeer the cell's mechanism for protein synthesis to produce the protein encoded by the gene.