Identification Of Structural Proteins From CSBV-LN And Structure Of Baculovirus Expression System

ObjectiveChinese sacbrood virus Liaoning strain (CSBV-LN) was purified and separated by SDS-PAGE, and the structural proteins were identified by mass spectrographic. In this base, the vector of VP1 gene was structured and the VP1 gene was expressed by the baculovirus expression system. This study is the basis for the further research of serodiagnosis kit of CSBV-LN.MethodsCSBV-LN was purified from diseased insects. The four main structural of CSBV were separated by SDS-PAGE. Structural proteins VP1, VP3 and VP0 were identified by mass spectrographic. The VP1 protein was cloned and sequenced to construct baculovirus expression vector, then transfected into sf9 insect cell. The protein which was secreted by sf9 insect cell was detected by Western-blotting.Results1. CSBV-LN virus prefiltration was got by purification.2. CSBV-LN was purified from diseased insects and was detected by SDS-PAGE from which structural proteins, VP1, VP3 and VP0, were identified after four proteins was analysed by mass spectrographic.3. The baculovirus expression system: VP1-pFastBacI was structured and the transposition was occurred in DH10Bac, and the recombinant vector: VP1-Bacmid was got.4. The recombination transposition vector was transfected into sf9 insect cell, and the diseased cells were collected to detect by Western-blotting. The objective band is about 31KD.Conclusions1. CSBV-LN was purified, and the virus prefiltration was high purity and high concentration.2. CSBV-LN was purified from diseased insects and was detected by SDS-PAGE from which structural proteins, VP1, VP3 and VP0, were identified after four proteins was analysed by mass spectrographic.3. The baculovirus expression system: VP1-pFastBacI was structured.4. The baculovirus expression vector of VP1 was structured and was expressed in the recombinant baculovirus, and a strain of recombinant baculovirus was get.