Studies On Expression Of HKLK1 In Pichia Pastoris

Human tissue kallikrein is a number of a closely related subfamily of serine proteinases, which specifically cleave peptide precursors to release potent vasodilative peptide kinin. Kininogen-Kallikrein system participates in regulation of local blood flow as well as the water and electrolyte balance in the kidney. Recent studies have suggested that kallikrein-kinin system is involved in control of human and animal hypertension. Moreover, the kallikrein gene expression may be related to the development of hypertension.The use of the methylotrophic yeast P Pastoris as a cellular host for the expression of recombinant proteins has become increasing popular in recent years. P. Pastoris is easier for genetic manipulation and culture than mammalian cells and can be grown to high cell densities. Equally important is that P. Pastoris, as an eukaryote expression system, can provide the potential for producing soluble, correctly folded recombinant proteins with correct post-translational modifications required for functionality. Additionally, linearized recombinant vectors can be inserted into the host genome at high frequency via homologous recombination to generate stable cell lines with high expression levels. A further benefit of the P. Pastoris system is that strong promoter is available to drive the expression of foreign gene(s) of interest, thus enabling production of large amounts of the target protein(s) with relatively easy techniques and low cost.In this research the coding region of human tissue prokallikrein was cloned into P. Pastoris expression vector pPIC9K and the Sal I- or Bgl II-linearized recombinant...