DNA construct
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] These elements can be as small as a few thousand base pairs (kbp) of DNA carrying a single gene, or as large as hundreds of kbp for large-scale genomic studies. 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. A DNA construct may express wildtype protein, prevent the expression of certain genes by expressing competitors or inhibitors, or express mutant proteins, such as deletion mutations or missense mutations. It can also prevent the expression of certain genes by encoding sequences of protein competitors or inhibitors. DNA constructs are widely adapted in molecular biology research for techniques such as DNA sequencing, protein expression, and RNA studies.
Typically, the vectors used in DNA constructs contain an origin of replication, a multiple cloning site, and a selectable marker.[2] Certain vectors can carry additional regulatory elements based on the expression system involved.
History
The first standardized vector, pBR220, was designed in 1977 by researchers in Herbert Boyer’s lab. The plasmid contains various restriction enzyme sites and a stable antibiotic-resistance gene free from transposon activities[3].
In 1982, Jeffrey Vieira and Joachim Messing described the development of M13mp7-derived pUC vectors that consist of a multiple cloning site and allow for more efficient sequencing and cloning using a set of universal M13 primers. Three years later, the currently popular pUC19 plasmid was engineered by the same scientists[4].
See also
References
- ^ Pinkert, Carl (2014). Transgenic animal technology: A laboratory handbook. Amsterdam: Elsevier. p. 692. ISBN 9780124095366.
- ^ Carter, Matt; Shieh, Jennifer C. (2010), "Molecular Cloning and Recombinant DNA Technology", Guide to Research Techniques in Neuroscience, Elsevier, pp. 207–227, doi:10.1016/b978-0-12-374849-2.00009-4, ISBN 978-0-12-374849-2, retrieved 2020-10-24
- ^ Bolivar, Francisco; Rodriguez, Raymond L.; Betlach, Mary C.; Boyer, Herbert W. (1977-11-01). "Construction and characterization of new cloning vehicles I. Ampicillin-resistant derivatives of the plasmid pMB9". Gene. 2 (2): 75–93. doi:10.1016/0378-1119(77)90074-9. ISSN 0378-1119.
- ^ Yanisch-Perron, Celeste; Vieira, Jeffrey; Messing, Joachim (1985-01-01). "Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mpl8 and pUC19 vectors". Gene. 33 (1): 103–119. doi:10.1016/0378-1119(85)90120-9. ISSN 0378-1119.
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