Gauthier L, Cousserans F, Veyrunes J C, Bergoin M
Proteine Performance S. A., Route d'Alès, Saint Christol lez Alès, France.
Virology. 1995 Apr 20;208(2):427-36. doi: 10.1006/viro.1995.1173.
We have cloned and sequenced a 1.7-kbp DNA fragment of the MmEPV genome encompassing the major polypeptide of the spindle-shaped inclusions gene termed fusolin. The sequence contained a single open reading frame of 1203 nt capable of coding for a polypeptide of 45.8 kDa. The 13 N-terminal amino acid (aa) residues were hydrophobic and could act as a signal peptide. The aa sequence also contained 13 cysteine residues very likely involved in paracrystal formation. This sequence showed significant homologies with the fusolins of two lepidopteran EPVs, the Choristoneura biennis EPV (CbEPV) and the Heliothis armigera EPV, and also with the 37K glycoproteins of Autographa californica and Orgyia pseudotsugata baculoviruses. No homology was found between the MmEPV fusolin and the 100K MmEPV spherulin, nor with the 110K polypeptide of the CbEPV and Amsacta moorei EPV spheroidins. These data were confirmed by Western blot analysis. Transfection of vaccinia-infected mammalian cells with a plasmid encompassing the fusolin sequence plus the upstream regulatory region resulted in transient expression of the gene. This indicated that the vaccinia transcription machinery is able to transcribe the fusolin gene. The fusolin was also expressed in insect cells via a recombinant baculovirus.
我们克隆并测序了苜蓿银纹夜蛾核型多角体病毒（MmEPV）基因组的一个1.7kbp的DNA片段，该片段包含纺锤形内含体基因的主要多肽，称为fusolin。该序列包含一个1203nt的单一开放阅读框，能够编码一个45.8kDa的多肽。其N端的13个氨基酸残基具有疏水性，可能作为信号肽。该氨基酸序列还包含13个半胱氨酸残基，很可能参与副晶体的形成。该序列与两种鳞翅目昆虫痘病毒（Choristoneura biennis EPV，CbEPV和棉铃虫EPV）的fusolin以及苜蓿银纹夜蛾核型多角体病毒和云杉毒蛾杆状病毒的37K糖蛋白具有显著的同源性。在MmEPV fusolin与100K MmEPV球形体蛋白之间，以及与CbEPV和摩尔粘虫EPV球形体蛋白的110K多肽之间未发现同源性。这些数据通过蛋白质免疫印迹分析得到证实。用包含fusolin序列加上游调控区的质粒转染痘苗病毒感染的哺乳动物细胞，导致该基因的瞬时表达。这表明痘苗病毒转录机制能够转录fusolin基因。fusolin也通过重组杆状病毒在昆虫细胞中表达。
