Cloning and expression of truncated form of tissue plasminogen activator in Leishmania tarentolae.
An expression cassette containing kringle 2 and serine protease domains (K2S), tissue plasminogen activator (tPA), together with a signal sequence derived from Leishmania tarentolae and two fragments of the small subunit ribosomal RNA locus, was introduced into L. tarentolae. The transfected cells produced recombinant K2S (rK2S) protein extracellularly with serine protease activity. Expression and enzyme activity of rK2S in the supernatant was 930 i.u./ml. The specific activity of purified rK2S was 7.4 U/mg of protein. Replacement of the human signal sequence tPA with the signal sequence derived from Leishmania increased the secretion of recombinant protein up to 30 times.
Department of Microbiology, Qom Branch, Islamic Azad University, 15 Khordad BLVD, Qom, Iran. nazari1102002@yahoo.com
Razieh
R
Nazari
Noushin
N
Davoudi
D006801:Humans; D018082:Kringles / Q000235:genetics; D007891:Leishmania / Q000235:genetics / Q000378:metabolism*; D008957:Models, Genetic; D012335:RNA, Ribosomal / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D057057:Serine Proteases / Q000235:genetics / Q000378:metabolism; D010959:Tissue Plasminogen Activator / Q000235:genetics / Q000378:metabolism*
D012335:RNA, Ribosomal; D011994:Recombinant Proteins; D057057:Serine Proteases; D010959:Tissue Plasminogen Activator