VACCINIA VIRUS SERPINS B13R (SPI-2) AND B22R (SPI-1) ENCODE M(R) 38.5K AND 40K, INTRACELLULAR POLYPEPTIDES THAT DO NOT AFFECT VIRUS VIRULENCE IN A MURINE INTRANASAL MODEL
VACCINIA VIRUS SERPINS B13R (SPI-2) AND B22R (SPI-1) ENCODE M(R) 38.5K AND 40K, INTRACELLULAR POLYPEPTIDES THAT DO NOT AFFECT VIRUS VIRULENCE IN A MURINE INTRANASAL MODEL
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DOI：
10.1016/s0042-6822(95)80028-x
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发表时间：
1995-01-10
期刊：
影响因子：
3.7
通讯作者：
SMITH, GL
中科院分区：
文献类型：
--
作者：
KETTLE, S;BLAKE, NW;SMITH, GL
对痘苗病毒西部储备株(WR)的B13R(SPI-2)和B22R(SPI-1)基因进行了鉴定。这些基因在感染早期被转录，预测的编码蛋白与丝氨酸蛋白酶抑制物(Serpins)超家族相似。通过引物延伸实验，将每个基因的5‘转录起始点分别定位于B13R和B22R开放阅读框(ORF)上游的71-72和31个核苷酸。每个ORF都在大肠杆菌中表达，并针对所产生的蛋白制备特异性抗血清。这些抗血清用于鉴定痘苗病毒感染细胞中B13R和B22R编码的蛋白分别为M(R)38.5K和M(R)40K的稳定、胞内和非糖基化蛋白。在所有测试的正痘病毒中都检测到B22R基因产物，包括牛痘、兔痘和牛痘病毒株WR、哥本哈根、塔什干、田坛、利斯特、惠氏、IHD-J和IHD-W。相比之下，B13R基因产物的分布较为有限，在哥本哈根、塔什干、利斯特和天坛没有检测到。通过瞬时显性选择，构建了仅缺失B13R或B22R编码序列(Delta B13R和Delta B22R)的活病毒缺失突变体和缺失基因恢复的突变株。缺失突变体在细胞培养中的生长与野生型病毒几乎没有区别。此外，在小鼠鼻腔模型中，每个缺失突变体的毒力与野生型和突变型病毒无法区分。(C)1995年学术出版社。
A characterization of genes B13R (SPI-2) and B22R (SPI-1) from vaccinia virus strain Western Reserve (WR) is presented. These genes are transcribed early during infection and the predicted encoded proteins show similarity to the superfamily of serine protease inhibitors (serpins). The 5' transcriptional initiation site of each gene was mapped by primer extension experiments to 71-72 and 31 nucleotides upstream of the B13R and B22R open reading frames (ORFs), respectively. Each ORF was expressed in Escherichia coli and specific antisera were raised against the protein produced. These antisera were used to identify the B13R- and B22R-encoded proteins in vaccinia virus-infected cells as stable, intracellular, nonglycosylated proteins of M(r) 38.5K and M(r) 40K, respectively. The B22R gene product was detected in all orthopoxviruses tested including cowpox, rabbitpox, and vaccinia strains WR, Copenhagen, Tashkent, Tian Tan, Lister, Wyeth, IHD-J, and IHD-W. In contrast, the B13R gene product had a more limited distribution and was not detected in Copenhagen, Tashkent, Lister, and Tian Tan. Viable virus deletion mutants that lacked only B13R or B22R coding sequences (Delta B13R and Delta B22R) and revertant viruses in which the deleted gene was restored were constructed by transient dominant selection. The growth of the deletion mutants in cell culture was indistinguishable from that of wild-type virus. Additionally the virulence of each deletion mutant was indistinguishable from wild-type and revertant viruses in a murine intranasal model. (C) 1995 Academic Press, Inc.