Designing and Expression of Bovine Chimeric Lactoferrampine - Lactoferricin Peptide with Aim of Recombinant Protein Production in Tobacco
Lactoferrin, which consists of lactoferampin and lactoferricin peptides, is one of the milk glycoproteins with anti-microbial properties. Lactoferrin is known as a component of intrinsic immunity which points to first barrier against microbes. The aim of this study was the chimeric synthesis of this two peptides and also producing transgenic product. One of the desired methods to produce recombinant peptides is by transferring their genes to plant cells and producing hairy roots. In this study, the sequence of chimeric Lfampin-Lfcin was codon optimized based on Tobacco expression system. The target sequence was artificially synthesized in pGH vector and then sub-cloned into the pBI121 expression vector. The expression vector was transferred to Agrobacterium rhizogenes through freez- defreezing process. Then it was transferred to Tobacco plant using leaf disc method. Extraction of recombinant protein was done using Tris-HCl method and purification process was accomplished by chromatography Ni-agarose columns. Finally, the results showed that the amount of recombinant protein was 1.52 mg/ml and its quality was also desirable. The results also showed that chimeric composition has a good inhibitory effect on pathogenic microorganisms.
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