Expression Of 45W-4B Gene Of Taenia Solium In E.coli And BmNPV-Bm System And Study On The Structure And Function Of 45W Protein

45W gene family of Taenia solium constitutes of a group of genes that have familiarstructure and function. Three types of 45W genes of T. solium, A type, B type and C typewere produced through alternative splicing. A transcript is composed of the four exonsⅠ, Ⅱ, Ⅲ and Ⅳ, B transcript is lack of exon Ⅱ, and C transcript is lack of exons Ⅱand Ⅲ. The same exon between different genes is highly homologous, but shows somevariations. 45W genes of T. solium were identified to be a talent candidate gene for generecombinant vaccine. In this study, 4B full-length and 3′truncated gene were successfullycloned into pET-28a vector, and expressed in BL21. The products of expression weredetected by Western blot with serum of human infected with Cysticercus Cellulosae, andshowed its antigenicity. The form of alternative splicing of 45W gene family interested many parasitologists.In these transcripts, 4B gene is a special one with the most variation contrast with theother genes of 45W. In this study, 4B protein was isolated and used to vaccinateintraperitoneally the Kunming line mice for twice, then the anti-serum of the mice waspooled for the following Western blot with the tissue protein of Cysticercus cellulosae.The Western blot appears the band in the 50 kDa, which shows that 4B protein fused with6 HIS have antigenicity, and can stimulate mice to produce antigen-specific antibody. Onthe other hand, it can be ascertained that 4B protein is presented in the tissue ofcysticercus cellulosae with the polymer. 4B gene and protein sequence is analyzed usingthe DNAstar software, and blasted using Protein Data base (PDB), showed that 4B proteinhave potential antigenicity, and in this protein there is a Fibronectin (FN) domain. It ispredicted that the 45W protein is related to forming of cysticercus.　 Expression system of BmNPV-Bm is high effective for producing recombinantprotein approach to its nature structure by the correct post-transcript process andmodification. 45W-4B gene of T. solium was cloned into baculovirus transfer vector ofpVL1393 and cotransfected into Bombyx mori nuclear polyhedrosis virus (BmNPV). Therecombinant BmNPV was screened, and used to infect Bm-N cells and silkworm. 4Bprotein was detected in the cell supernatant and haemolyph by Western blot using serumof human (and swine) infected with Cysticercus Cellulosae. Successful expression of 4Brecombinant protein in BmNPV-Bm will be significant for study the structure and functionof 4B protein, and development of the effective recombinant vaccine against T. solium andCysticercus Cellulosae.