Construction Of Bacterial Artificial Chromosome Vectors Containing Human Tissue-type Plasminogen Activator Mutant Gene Specially Expressed In Mammary Gland With Red Recombination System

Tissue-type plasminogen activator has immense clinical significance as thrombolytic agents for management of thromboembolism such as myocardial infarction. In this study we use a variant form of human Tissue-type plasminogen activator (t-PAm) with prolonged clearance and enhanced activity. The high cost of production of t-PAm through large-scale culture in mammalian cell lines which is the main way at present leads to the high price of this medicine.Transgenic animal as mammary gland bioreactors are an attractive production system for high productivity of pharmaceutically useful proteins. It has many advantages prior to other production systems. All above animal cells can synthesize proteins with the appropriate post-translational modifications. But a major problem in the production of transgenic animals as mammary gland bioreactors is the position effect leading to the silence of the expression. Given the high capacity of bacterial artificial chromosomes, using BAC as transgenic vectors may be an ideal tool because they can contain all major important elements of lacto protein. Some of these elements work as insulator overcoming the position effects. The transgenic techniques via BAC vectors may be another nice approach to obtain transgenic animals as mammary gland bioreactors with high efficiency and high expression except for gene targeting.In our study, we successfully constructed a BAC vector containing t-PAm gene which is regulated by P-casein locus. First, we rapidly modified bacterial artificial chromosome containing mouse P-casein gene by homologous recombination...