Pectin is polysaccharides that are usually found in the plant cell walls. Pectin is involved in many functions such as plant growth, intercellular interactions, cell wall structure, signal transduction, cell expansion, pollen tube development and fruit maturation and so on (Claudia et al, 2018). Many bacterial organisms produce enzymes that break down polysaccharide networks to facilitate host tissue invasion. Pectate lyase is one of the enzymes. This enzyme stimulates the separation of pectin by separating glycosidic bonds between monosaccharide residues (Scavetta et al, 1999). Therefore, this enzyme has been used in the food and winemaking industry for the processing of fruit juices (Benjamin, 2012).
pET-28a plasmid has MCS (multiple cloning site) which contains various restriction sites.
Restriction enzymes digest MCS and cloned genes. This process produces sticky ends of the plasmid vector and target gene allowing ligation of them.
Benjamin, V. (2012). Food industrial processes: Methods and Equipment. Books of Demand. pp195
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Claudia, L, E., Elizabeth, C, M., René, B, Q., Yolanda, L, F., and Agustín, R, C. (2018). Pectin and Pectin-Based Composite Materials: Beyond Food Texture. [online] Ncbi.nlm.nih.gov. Available at: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6017442/ [Accessed 1 Feb. 2020].
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Scavetta, RD., Herron, SR., Hotchkiss, AT., Kita, N., Keen, NT., Benen, JA., Kester, HC., Visser, J. and Jurnak, F. (1999). Structure of a plant cell wall fragment complexed to pectate lyase C. [online] Ncbi.nlm.nih.gov. Available at: https://www.ncbi.nlm.nih.gov/pubmed/10368179 [Accessed 1 Feb. 2020].