Kato A, Fujino M, Nakamura T, Ishihama A, Otaki Y
Nippon Institute for Biological Science, Tokyo, Japan.
Virology. 1995 Jun 1;209(2):480-8. doi: 10.1006/viro.1995.1280.
The genomic DNA of chicken anemia virus (CAV) was cloned and sequenced from a Japanese isolate CAA82-2. The nucleotide sequence of CAA82-2 isolate was 98% identical with that of the European Cuxhaven-1 strain (Noteborn et al., J. Virol. 65, 3131-3139, 1991). Nine open reading frames (ORFs) consisting of more than 100 nucleotides were found, i.e., four ORFs (CA1-CA4) on the plus strand and five ORFs (CA1R-CA5R) on the minus strand. These ORFs with the exception of CA4 are conserved between the two CAV isolates. All of these ORFs were expressed in Escherichia coli as fusion proteins with beta-galactosidase. By Western blot analysis, the CA2 and CA3 fusion proteins were found to react with CAV-infected chicken sera. Rabbit hyperimmune sera against the CA1, CA2, and CA3 fusion proteins were produced and tested their reactivity to CAV-infected cells. Two viral proteins with the apparent size of 54 and 16 kDa reacted with the antibodies against CA1 and CA3 fusion proteins, respectively. The 16-kDa protein, CA3, was suggested to be a major immunogen on CAV infection.
从日本分离株CAA82 - 2中克隆并测序了鸡贫血病毒（CAV）的基因组DNA。CAA82 - 2分离株的核苷酸序列与欧洲库克斯港 - 1株的核苷酸序列有98%的同一性（Noteborn等人，《病毒学杂志》65，3131 - 3139，1991）。发现了九个由100多个核苷酸组成的开放阅读框（ORF），即正链上的四个ORF（CA1 - CA4）和负链上的五个ORF（CA1R - CA5R）。除CA4外，这些ORF在两种CAV分离株之间是保守的。所有这些ORF都在大肠杆菌中作为与β - 半乳糖苷酶的融合蛋白表达。通过蛋白质免疫印迹分析，发现CA2和CA3融合蛋白与CAV感染的鸡血清发生反应。制备了针对CA1、CA2和CA3融合蛋白的兔超免疫血清，并检测它们对CAV感染细胞的反应性。两种表观大小分别为54 kDa和16 kDa的病毒蛋白分别与针对CA1和CA3融合蛋白的抗体发生反应。16 kDa的蛋白CA3被认为是CAV感染的主要免疫原。
