Snails Blood Ceruloplasmin Cdna Cloning And Immunological Properties

A rapid and simple blood serum diagnosing method has been set up by using Oncomelania hupensis hemocyain(ohH).We have prepared different antiserum from different developmental stages of S. japonicum (Cercaria, Schistosomulum, Adult worm and Egg), and have made research into the interaction between antibody and antigen by a method of ELISA using purified Oncomelania hupensis hemocyain as antigen, antiserum as first antibody. In this study, it shows that ohH shares carbohydrate epitopes with different developmental stages of Schistosoma. It is expected to be a candidate for vaccine against S. japonicum.By the methods of RT-PCR and 3'RACE , designing primers by CODEHOP, We have cloned seven cDNA segments, including 1.563kb, 1.148kb, 2.414kb,2.205kb,1.796kb ,2.425kb and 3.174kb. By using the SeqMan, we get a 9.648kb ,a long segment by joining seven small segments together. We blast seven cDNA segments on the web of NCBI, it shows that the seven Cdna segments all have higher homology by contrast with cDNAs for hemocynion from other animals in mollusks which have the highest homology with Aplysia Californica mRNA for hemocynion. Compared with the equivalent cDNAs for hemocynion from other mollusks, we find they have higher homology, which the highest is up to 72%. We translate 9.648kb segment into animo acid by Primer Premier5.0 from the third base, and it shows that it has 65% homology comparing with equivalent animo acid for hemocynion from other mollusks. On the whole, it is fully convinced that all the cloned cDNA segments are ohH.We contrast 3313aa amino partial sequence with other mollusks amino sequence for hemocyanin from the GenBank of NCBI and build a cladogram by choosing 15 typical mollusks hemocyanin amino sequence. It shows that they have closest affinity between Oncomelania hupensis hemocyanin and Aplysia Californica hemocynion which is consistent with the results from cDNA alignment. They are the first time cloned by us in the world, and lay foundations for the latter research of structure and function of ohH.