Selected topics of multidisciplinary science Biotechnology
Biotechnology is a multidisciplinary science that covers different techniques and processes, in addition to the great advances it has had in recent years, molecular biology has opened the door to obtain new organisms and design proteins (CAR/PL, 2003). The development of new molecular tools, whose use has increased significantly since the discovery of the PCR, cloning, sequencing, fluorescence detection technology and accessibility to a large amount of information on the web (Palomino & González, 2014). Bacteria and other organisms contain small double strand DNA DNA DNA molecules, called “plasmids” that contains a gene that provides resistance to antibiotics, in addition to having multi -cloning sites and unique cutting site which makes themThe most used cloning vectors (Zapata, 2004).
For the extensive knowledge about the genetics of bacteria and the aforementioned the main model used as an expression system is Escherichia coli. To exploit this ability it is necessary. coli (Galván-Cejudo, to. et al. s/f). The purification of the plasmid by means of the Miniprep method, to isolate the plasma DNA of the rest of the bacterial cellular components by the alkaline lysis method, where the cells are broken by an alkaline shock (NaOH) and a detergent (dodecyl sodium sulfate;Sds) and the plasmid with ethanol is precipitated (Damián, P. et al. 2013).
Endonucleases are restriction enzymes that cut DNA large molecules into specific places, generating smaller DNA fragments with “cohesive” or shaved ends and the enzyme ligase that generates covalent unions between DNA fragments (Zapata, 2004), this informationIt is of great importance in the production process of a homemade ladder. Adding the electrophoresis technique that is based on the study of the movement of molecules loaded in an electric field and is particularly used for the characterization and analysis of loaded biological polymers, such as proteins and nucleic acids (Damián, P. et al. 2013), within agarose or polyacrylamide gels in concentration percentages to increase or decrease permeability in order to separate samples by molecular weight.
Finally, a very important analytical technique used to detect the expression of a specific protein in a crude extract, through the use of antibodies is the Western Blot (Advansta, 2009). It implies the separation by electrophoresis by the SDS-Page system and a quantitative and irreversible transfer to a nitrocellulose membrane of the proteins of a complex mixture (total cell phone). The antigens that have been transferred to the membrane are recognized by specific monoclonal or polyclonal antibodies (from the 2007 source).
Leave feedback