Isolation And Identification Of Vibrio Parahaemolyticus Of Scylla Serrata And Preparation Of Monoclonal Antibody Against Thermolabile Hemomysin

Scylla serrata is one species of Scylla,Portunidae,Brachyra,Decapoda,which is one of the main edible crabs in China.Recently,a disease had caused the mass death of S.serrata on the south-east coast of China,resulting in great loss in aquatic products.Vibrio Parahaemolyticus (VP)is one of the key microbes who made S.serrata sick.In this study,strain S,was isolated successfully by steak plate method.Strain S was identified primarily as VP by the morphological characteristic and the physiological and biochemical properties of bacteria.Gene tlh of VP was obtained by PCR,then the expression vector pET32a(+)which contained tlh gene was transformed into E.coli(BL21)for expression of thermolabile hemomysin(TLH)in LB medium by inducing with IPTG.The expressed TLH was used to immunize mouse after purification.Titer of mouse serum was detected by enzyme linked immonosorbant assay(ELISA).A hybridoma cell that could stably secret anti-TLH monoclonal antibody was obtained through cell fusion method,and named CH-112.