Yao B, Pang Y, Fan Y, Zhao R, Yang Y, Wang T
Biotechnology Research Center, Chinese Academy of Agricultural Sciences, Beijing, China.
Sci China C Life Sci. 1996 Apr;39(2):199-206.
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.
考虑到影响基因转录、翻译以及mRNA稳定性的因素，在不改变所编码多肽氨基酸组成的情况下，根据杆状病毒多角体蛋白基因以及植物基因密码子的使用偏好、总体G+C含量和密码子第三位碱基的G+C含量，设计并合成了编码昆虫特异性神经毒素的AaIT基因。将AaIT基因融合在合成的gp67信号序列之后，然后通过转移载体pSXIV VI+X3重组到粉纹夜蛾核多角体病毒（TnNPV）的基因组中。筛选出重组病毒TnNPV-AaIT（occ+-gal-）。Southern杂交和SDS-PAGE结果表明，AaIT基因已整合到病毒基因组并表达。对三龄粉纹夜蛾幼虫的生物测定表明，重组病毒TnNPV-AaIT能够缩短杀虫时间并提高对重要农业害虫的杀虫效果。
