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Cloning and expression of the human tissue kallikrein 1 and 6

Grant number: 07/04339-1
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): April 01, 2008
Effective date (End): December 31, 2008
Field of knowledge:Biological Sciences - Biochemistry - Molecular Biology
Principal Investigator:Luciano Puzer
Grantee:Priscila Tomie Leme Ike
Home Institution: Centro de Ciências Biológicas e da Saúde (CCBS). Universidade Federal de São Carlos (UFSCAR). São Carlos , SP, Brazil

Abstract

Human tissue kallikreins 1 (hK1) and 6 (hK6) belong to a 15-member family of closely related serine proteases. hK1 is mainly known as the enzyme that hydrolysis the low molecular weight kininogen to generate lysil-bradykinin. There is not much information about the hK7 physiological roles, but this enzyme appears to be implicated in the sclerosis multiple development. In this project we are going to cloning and to express the hK1 and hK6. The genes KLK1 and KLK6 will be amplified from a cDNA panel of multiple tissues, sub-cloned in the expression vector pET28a(+), and transformed in the E. coli cells (rosetta). The enzymes expressed in this system will be purified using nickel columns, and the identification we be done by N-terminal sequence and molecular weight of the enzymes. In the further, both enzymes hK1 and hK6 we be used to produce antibody.