MOLECULAR-CLONING OF DNA COMPLEMENTARY TO MESSENGER-RNA OF THE BACULOVIRUS AUTOGRAPHA-CALIFORNICA NUCLEAR POLYHEDROSIS-VIRUS - LOCATION AND GENE-PRODUCTS OF RNA TRANSCRIPTS FOUND LATE IN INFECTION
MOLECULAR-CLONING OF DNA COMPLEMENTARY TO MESSENGER-RNA OF THE BACULOVIRUS AUTOGRAPHA-CALIFORNICA NUCLEAR POLYHEDROSIS-VIRUS - LOCATION AND GENE-PRODUCTS OF RNA TRANSCRIPTS FOUND LATE IN INFECTION
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DOI：
10.1128/jvi.44.3.782-793.1982
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发表时间：
1982-01-01
影响因子：
5.4
通讯作者：
MILLER, LK
中科院分区：
文献类型：
--
作者：
ADANG, MJ;MILLER, LK
通过反转录合成与晚期加州核型多角体病毒（AcNPV）mRNA互补的DNA，并使用pBR322作为载体克隆到大肠杆菌中。在筛选 45 个 AcNPV 同源克隆时，区分出了 11 种不同的 c[互补]DNA。 cDNA 同源区域相对于 AcNPV 物理图谱的定位表明，11 个 cDNA 分散在整个基因组中。最丰富的 cDNA 插入，代表 .apprx。 1/3 的晚期病毒 mRNA 与 AcNPV HindIII-P、Q 和 EcoRI-P 片段同源。在线性化 AcNPV 物理图上，该区域的转录方向是从左到右。杂交选择和体外翻译表明该区域编码 7200 道尔顿 (7.2K) 蛋白质，该蛋白质与细胞外非封闭形式病毒 (NOV) 中发现的次要蛋白质共迁移。多角体蛋白基因（封闭病毒形式的主要结构蛋白）至少部分位于 AcNPV 图谱的 HindIII-V/EcoRI-I 区域。多角体蛋白转录物代表.apprx。感染后 27 小时，1/4 的病毒含 Poly(A) RNA。另一个相对丰富的cDNA与HindIII-A/EcoRI-C/SstI-G区域同源，并且由该cDNA选择的RNA指导2种蛋白质（31K和30K）的合成。鉴定了另外 5 种 cDNA 选择的 RNA 的蛋白质产物。 AcNPV L-1 基因组的 HindIII-D/EcoRI-O、HindIII-C/EcoRI-D、HindIII-B1/EcoRI-E 和 HindIII-B2/EcoRI-H 区域与 RNA 同源，分别指导 57K 蛋白质、25K 蛋白质、61K 蛋白质和 37K 蛋白质（加上少量 26K 蛋白质）的合成。由与 AcNPV 图谱的 HindIII-P/EcoRI-B 区域同源的 cDNA 选择的晚期 mRNA 指导 31K 和 30K 蛋白质的合成，这些蛋白质与从 HinDIII-A/EcoRI-C/SstI-G cDNA 选择的 RNA 翻译的 31K 和 30K 蛋白质共迁移。其他三个 cDNA 尚未与特定蛋白质产物相关联。
DNA complementary to late A. californica nuclear polyhedrosis virus (AcNPV) mRNA were synthesized by reverse transcription and cloned in Escherichia coli by using pBR322 as a vector. Eleven different c[complementary]DNA were distinguished in the screening of 45 AcNPV-homologous clones. Location of the regions of cDNA homology with respect to the AcNPV physical map showed that the 11 cDNA were dispersed throughout the genome. The most abundant cDNA insertion, representing .apprx. 1/3 of the late viral mRNA, was homologous to the AcNPV HindIII-P,Q and EcoRI-P fragments. The direction of transcription in this region was from left to right on a linearized AcNPV physical map. Hybridization selection followed by in vitro translation showed that this region encoded a 7200-dalton (7.2K) protein which comigrated with a minor protein found in the extracellular nonoccluded form of the virus (NOV). The gene for polyhedrin, the major structural protein of the occluded virus form, was located, at least in part, in the HindIII-V/EcoRI-I region of the AcNPV map. The polyhedrin transcript represented .apprx. 1/4 of the viral poly(A)-containing RNA at 27 h postinfection. Another relatively abundant cDNA was homologous to the HindIII-A/EcoRI-C/SstI-G region, and RNA selected by this cDNA directed the synthesis of 2 proteins (31K and 30K). The protein products of 5 other cDNA-selected RNA were identified. The HindIII-D/EcoRI-O, HindIII-C/EcoRI-D, HindIII-B1/EcoRI-E and HindIII-B2/EcoRI-H regions of the AcNPV L-1 genome were homologous to RNA which directed the synthesis of a 57K protien, a 25K protein, a 61K protein and a 37K protein (plus a minor 26K protein), respectively. Late mRNA selected by a cDNA homologous to the HindIII-P/EcoRI-B region of the AcNPV map directed the synthesis of 31K and 30K proteins which comigrated with the 31K and 30K proteins translated from RNA selected by the HinDIII-A/EcoRI-C/SstI-G cDNA. Three other cDNA have not been correlated yet with specific protein products.