White spot disease caused by white spot syndrome virus (WSSV) leads to a devastating loss in shrimp farming. Up to date there is no effective way to prevent the attack by WSSV to shrimps. The WSSV viral envelop consists of at least 35 different proteins, of which VP28 and VP26 are the most abundant, accounting for approximately 60 percent of the envelope. The WSSV envelope protein VP28 has been found to playa key role in viral attachment and penetration into shrimp cells. This protein is highly antigenic and can stimulate the immune response in shrimps and that is why this envelop protein has been used as target protein in WSSV diagnostics and vaccine production to protect shrimps against the WSSV. In this study, VP28 ,encoding gene was amplified by PCR and cloned into pDG364-cotB vector to form a recombinant construct pDG364-cotB VP28. The targeted gene cotB-vp28 from pDG364-cotB-VP28 was then successfully integrated into the chromosome of B. subtilis PY79 in the form of CotB-VP28, distrupting the amylase encoding gene of the host. The obtained colonies lost their starch hydrolyzing activity and the Westren blotting analysis data showed the presence of 63 kDa protein band using polyclonal antibodies against VP28. These results confirmedthe successful expression of CotB-VP28 on the recombinant Bacillus subtilis spores. The expression of CotBVP28 on the spore surface was also detected by immunofluorescence analysis. The improvement of VP28 expression and evaluation of immune response and protection of shrimps by the recombinant Bacillus subtilis spores against WSSV are in progress.
Lọc theo danh mục
Lượt truy cập :
17,525,229
Xem toàn văn