Theilmann D A, Summers M D
Department of Biology, Texas A&M University, College Station 77843-2475.
Virology. 1988 Dec;167(2):329-41.
The Campoletis sonorensis virus (CsV; Polydnaviridae) genome consists of at least 28 closed circular superhelical (SH) DNAs. In this study we used complete clones of four SH DNAs to analyze viral transcription both in the adult parasitic wasp host Campoletis sonorensis (Ichneumonidae) and in the lepidopteran host, Heliothis virescens (Noctuidae). CsV genes are expressed in parasitized H. virescens, but no viral transcripts had been characterized from C. sonorensis until this study. The clones of the SH DNAs B, H, M, and O1 were used to probe Northern blots of poly(A)+ RNA isolated from C. sonorensis reproductive tissue and from parasitized H. virescens larvae. All four SH DNAs hybridized to viral transcripts. SH-H,-M, and -O1 hybridized to messages expressed in both hosts. SH-B and -M hybridized to transcripts that were detected only in either C. sonorensis reproductive tissue or parasitized H. virescens larvae. These results suggest that some CsV genes are expressed in a host-specific manner. In a previous study we identified a family of imperfectly conserved tandemly repeated 540-bp repeat elements on SH-B,-H and -O1 (D. A. Theilmann and M. D. Summers, 1987 J. Virol. 61; 2589-2598). Hybridization of the 540-bp repeat regions to Northern blots showed that they were all homologous to viral transcripts. A cDNA clone of a mRNA that is transcribed from the 540-bp repeat region of SH-B was isolated from a lambda gt 10 library and completely sequenced. The sequence data revealed that the 540-bp repeat element was contained within the open reading frame of this gene. These results indicate that transcribed sequences homologous to the 540-bp repeat elements represent a second gene family to be identified within the CsV genome.
Campoletis sonorensis病毒(CsV;多DNA病毒科)基因组由至少28个闭环超螺旋(SH)DNA组成。在本研究中,我们使用了四个SH DNA的完整克隆,来分析病毒转录情况,研究对象包括成年寄生黄蜂宿主 Campoletis sonorensis(姬蜂科)和鳞翅目宿主烟芽夜蛾(夜蛾科)。CsV基因在被寄生的烟芽夜蛾中表达,但在本研究之前,尚未从 Campoletis sonorensis中鉴定出病毒转录本。SH DNA的B、H、M和O1克隆用于探测从 Campoletis sonorensis生殖组织和被寄生的烟芽夜蛾幼虫中分离的聚腺苷酸加尾(poly(A)+)RNA的Northern印迹。所有四个SH DNA都与病毒转录本杂交。SH-H、-M和-O1与在两个宿主中都表达的信息杂交。SH-B和-M与仅在 Campoletis sonorensis生殖组织或被寄生的烟芽夜蛾幼虫中检测到的转录本杂交。这些结果表明,一些CsV基因以宿主特异性方式表达。在先前的一项研究中,我们在SH-B、-H和-O1上鉴定了一个不完全保守的串联重复540 bp重复元件家族(D. A. Theilmann和M. D. Summers,1987年,《病毒学杂志》61卷;2589 - 2598页)。540 bp重复区域与Northern印迹的杂交表明,它们都与病毒转录本同源。从λgt 10文库中分离出一个从SH-B的540 bp重复区域转录的mRNA的cDNA克隆,并进行了全序列测定。序列数据显示,540 bp重复元件包含在该基因的开放阅读框内。这些结果表明,与540 bp重复元件同源的转录序列代表了CsV基因组中有待鉴定的第二个基因家族。