The enzyme is found in prokaryotes and eukaryotes with the same general function but in different structural forms suitable to the environment they function in. In E. coli, alkaline phosphatase is found in the periplasmic space while in humans is found in bile ducts, bone, intestine, placenta, and tumors.
Typical uses in the lab for alkaline phosphatases include: removing the phosphates groups on the 5' end to prevent the DNA from ligating (the 5' end attaching to the 3' end), thereby keeping DNA molecules linear until the next step of the process for which they are being prepared; also, removal of the phosphate groups allows radiolabeling (replacement by radioactive phosphate groups) in order to measure the presence of the labeled DNA through further steps in the process or experiment.
Figure 1: Alkaline phosphatase reaction.
The common reaction buffer used is the Alkaline phosphatase 10X Reaction Buffer (M183A).
Bibliography
·
Telega, Grzegorz
W.,Learn more about Alkaline phosphatase,2018. Disponível em: https://www.sciencedirect.com/topics/neuroscience/alkaline-phosphatase.
Acesso em 13 de novembro de 2018.
·
Sharma, Ujjawal; Pal, Deeksha;
Prasad, Rajendra; Alkaline Phosphatase: An Overview, 2014. Disponível em: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4062654/.
Acesso em 13 de novembro de 2018.· Phosphatase, Alkaline. Disponível em: http://www.worthington-biochem.com/bap/default.html. Acesso em 13 de novembro de 2018.
· Disponível em: https://www.promega.com/-/media/files/resources/protocols/product-information-sheets/g/alkaline-phosphatase-calf-intestinal-protocol.pdf. Acesso em 13 de novembro de 2018.
Ana Rita Pacheco (a83934) | Angelina Eiras (a80415) | Margarida Lima (a84229) | Maria Freitas (a84345) | Maria Rui Ferreira (a85952)
University of Minho | School of Sciences | Department of Biology
Degree in Aplicated Biology | Genes and Genomes
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