[Construction of a novel Bm NPV Bac to Bac system].
A Bi-Shuttle vector Bm-Bacmid was constructed by co-infecting Bm N cells with wild type genomic DNA from BmNPV and Ac-Bacmid DNA. It could not only replicate in E. coli cells as a large plasmid and but also remain infectious when induced into Bm N or Sf9 cells. Recombinant virus rBmHBe was obtained after transposition of a donor plasmid carrying Hepatitis Be antigen gene (HBeAg) into att Tn7, and was demonstrated by Southern blotting. SDS-PAGE analysis showed that HBeAg gene were highly expressed in Bm N cells. By ELISA testing, the highest antigenecity titer of HBeAg protein in cell cultural medium was up to a dilution of 1:32,000. Although HBeAg protein also presented in the Bm N cells the titer was only 1:2000. The HBcAg protein was much fewer than HBeAg (< 1:160) whatever in culture medium and in cells. The results showed that Bm N cells was able to recognize the signal peptide sequence and cut it correctly for HBeAg protein's excreting production.
Deng
X
X
Nanjing Command Military Medical Institute, Nanjing 210002.
Zhu
Y
Y
Diao
Z
Z
Qi
Y
Y
Zhou
Z
Z
D000818:Animals; D012831:Bombyx / Q000166:cytology / Q000378:metabolism; D002478:Cells, Cultured; D004251:DNA Transposable Elements; D004274:DNA, Recombinant / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D005822:Genetic Vectors; D006513:Hepatitis B e Antigens / Q000096:biosynthesis / Q000235:genetics*; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D010957:Plasmids; D018411:Spodoptera / Q000166:cytology / Q000378:metabolism; D014162:Transfection
D004251:DNA Transposable Elements; D004274:DNA, Recombinant; D004279:DNA, Viral; D006513:Hepatitis B e Antigens