The Clone And Expression Of Hemolysin Gene From Leptospira Interrogans Serovar Lai And The Primary Investigation Of Functions Of Hemlysins

Leptospirosis is a worldwide spirochetal zoonosis caused by pathogenic Leptospira. Patients affected by pathogenic Leptospira have an acute febrility and pathological alteration of organs. In some serious cases, the illness may be fatal to patients. Vascular endothelial lesions are characterized in the early stage of leptospirosis. The widespread petechial hemorrhages are apparent in various organs and tissues in man and animals. In the late stage, renal failure, liver cell necrosis, and jaundice have appeared.In this thesis, firstly, the genomic DNA of Leptospira interrogans serogroup Icterohaemorrhagiae serovar lai type strain #56601 was used as plate, eight pieces of hemolysin genes coding hemolysin proteins LA1029, LA4004, LA0327, LA3937, LA0378, LA1650, LA3050 and LA1027 were cloned by PCR method. Then 8 recombinant expression plasmids of the hemolysin proteins were constructed and successfully expressed in E.coli as a fusion protein. Afterward, the hemolytic activity of these fusion protein were assayed.Experiment results showed that all the 8 fusion proteins have clear hemolytic zones on sheep blood agar. Among the 8 proteins, LA1029, LA1027, LA4004, LA3050 showed sphingomyelinase...