Isolation, Characterization, Mapping and Expression analysis of Porcine Tissue kallikreins

Abstract

Kallikreins are members of a multigene family of serine proteases that are widespread throughout living organisms. They are found in diverse tissue specific patterns and are known to have highly diverse physiological functions such as fertilization, digestion, regulation of blood flow, blood coagulation, inflammatory responses, endothelial cell migration, tissue remodeling, tumor-cell invasion and programmed cell death as well as disease related specialized enzymatic activities. To gain insight about the structure and evolutionary origins of the kallikrein gene region in the pig, we have isolated several BAC (Bacterial Artificial Chromosome) clones containing members of the porcine tissue kallikrein gene family and constructed a BAC clone based Physical map of the porcine greater kallikrein gene region using hybridization based dot-blot analysis and DNA fingerprinting techniques. We have also mapped the kallikrein gene region in the porcine genome using radiation-hybrid mapping to chromosome 6q1.2. We have further isolated several porcine tissue kallikreins using RT-PCR and conserved sequence information from orthologous human kallikrein genes and have carried out quantitative real-time PCR based, expression analysis to evaluate the expression of tissue kallikreins in the porcine endometrium and conceptus during the estrous cycle and early embryonic development as a first step towards understanding the tissue expression of pig kallikrein gene family members and to better understand the role of kallikreins in placental development and embryonic survival in the pig.