Construction of an insecticidal baculovirus expressing insect-specific neurotoxin AaIT.
Considering the factors which affect gene transcription, translation and the stability of mRNA, without changing the amino acid composition of the encoded polypeptide, AaIT gene encoding insect-specific neurotoxin was designed and synthesized according to bias in codon choice, overall G+C content and G+C content of bases at the third position in codons of polyhedrin genes of baculovirus and of plant genes as well. AaIT gene was fused behind a synthetic gp67 signal sequence and then recombined into the genome of Trichoplusia ni nuclear polyhedrosis virus (TnNPV) by transfer vector pSXIV VI+X3. The recombinant virus TnNPV-AaIT (occ+-gal-) was screened. The results of Southern blotting and SDS-PAGE demonstrated that AaIT gene had integrated into the genome of virus and expressed. Bioassays on the 3rd-instar Trichoplusia ni larvae showed that recombinant viruses TnNPV-AaIT could shorten the time of killing insect and improve the efficacy of killing agronomically important insects.
Yao
B
B
Biotechnology Research Center, Chinese Academy of Agricultural Sciences, Beijing, China.
Pang
Y
Y
Fan
Y
Y
Zhao
R
R
Yang
Y
Y
Wang
T
T
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D004274:DNA, Recombinant; D015870:Gene Expression; D007313:Insecta / Q000821:virology*; D008969:Molecular Sequence Data; D009036:Moths / Q000821:virology*; D009498:Neurotoxins / Q000235:genetics*; D017924:Nucleopolyhedroviruses / Q000235:genetics; D010572:Pest Control, Biological; D012604:Scorpion Venoms / Q000235:genetics
D004274:DNA, Recombinant; D009498:Neurotoxins; D012604:Scorpion Venoms