Rodrigues J C, De Souza M L, O'Reilly D, Velloso L M, Pinedo F J, Razuck F B, Ribeiro B, Ribeiro B M
Embrapa Genetic Resources and Biotechnology, Brasília, Brasil.
Virus Genes. 2001 Jan;22(1):103-12. doi: 10.1023/a:1008142621359.
The Anticarsia gemmatalis nucelopolyhedrovirus (AgMNPV) egt gene was cloned, sequenced and its expression characterized by RT-PCR and western blot analysis. Sequence analysis of the gene indicated the presence of an open reading frame (ORF) of 1482 nucleotides, which codes for a polypeptide of 494 amino acids. ATATA box and a conserved regulatory sequence (CATT) found in other baculovirus early genes were present in the promoter region of the egt gene. A poly-A consensus sequence was present in the 3' untranslated region (3'-UTR) of the gene. Homology comparisons showed that the EGT protein of AgMNPV is most closely related (95.9% amino acid sequence identity) to the EGT from the Choristoneura fumiferana DEF nucleopolyhedrovirus (CfDEF). Transcriptional analysis of the AgMNPV egt gene showed that egt-specific transcripts can be detected both early and late in infection. The EGT protein was detected, by western blot analysis, in the intra- (from 12 to 48 h post-infection) and extra-cellular (from 12 to 96 h post-infection) fractions of infected insect cells. The AgMNPV Bgl II-F fragment, which has homology to the AcMNPV ie-1 gene, was cloned and used to cotransfect SF21 cells with the cloned AgMNPV egt gene. EGT activity was observed, suggesting that AgMNPV ie-1 can transactivate egt expression.
克隆了粉纹夜蛾核型多角体病毒（AgMNPV）的蜕皮甾体尿苷二磷酸葡萄糖基转移酶（egt）基因，进行了测序，并通过逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹分析对其表达进行了表征。该基因的序列分析表明存在一个1482个核苷酸的开放阅读框（ORF），其编码一个由494个氨基酸组成的多肽。在egt基因的启动子区域存在其他杆状病毒早期基因中发现的ATATA框和一个保守调控序列（CATT）。在该基因的3'非翻译区（3'-UTR）存在一个聚腺苷酸共有序列。同源性比较表明，AgMNPV的EGT蛋白与云杉色卷蛾DEF核型多角体病毒（CfDEF）的EGT关系最为密切（氨基酸序列同一性为95.9%）。AgMNPV egt基因的转录分析表明，在感染的早期和晚期均可检测到egt特异性转录本。通过蛋白质免疫印迹分析，在感染昆虫细胞的细胞内（感染后12至48小时）和细胞外（感染后12至96小时）部分检测到了EGT蛋白。克隆了与苜蓿银纹夜蛾核型多角体病毒（AcMNPV）ie-1基因具有同源性的AgMNPV Bgl II-F片段，并将其与克隆的AgMNPV egt基因共转染SF21细胞。观察到了EGT活性，表明AgMNPV ie-1可以反式激活egt表达。
