Makimura K, Peng F Y, Tsuji M, Hasegawa S, Kawai Y, Nonoyama M, Tanaka A
Department of Virology, Tampa Bay Research Institute, St. Petersburg, FL 33716.
Virus Genes. 1994 Jan;8(1):15-24. doi: 10.1007/BF01703598.
Polymerase chain reaction (PCR) to amplify MDV DNA and subsequent sequencing identified the junction of TRL/UL, UL/IRL, IRS/US, and US/TRS. The TRL/UL junction is located 192 bp downstream of the last EcoRI site in the TRL region, while the UL/IRL junction is located 192 bp upstream of the first EcoRI restriction enzyme site in the IRL region. The IRS/US junction is located 950 bp downstream of the second EcoRI site in the IRS region, while the US/TRS junction is located 950 bp upstream of the first EcoRI restriction enzyme site in the TRS region. BamHI restriction enzyme mapping of one of the PCR products identified two novel DNA subfragments, BamHI-U2 and -P4, upstream of the US/TRS junction of the MDV genome. Sequencing of the BamHI-D fragment revealed a novel open reading frame (ORF) encoding a 155 amino acid protein. The TRL/UL junction is located in this ORF. The N-terminal 65 amino acids of this protein is homologous to the N-terminal region of the previously reported pp38, which is located in the UL/IRL region. Computer-assisted analysis indicated that both are transmembrane proteins and that they share an antigenic domain.
通过聚合酶链反应（PCR）扩增MDV DNA并进行后续测序，确定了TRL/UL、UL/IRL、IRS/US和US/TRS的连接处。TRL/UL连接处位于TRL区域最后一个EcoRI位点下游192 bp处，而UL/IRL连接处位于IRL区域第一个EcoRI限制性酶切位点上游192 bp处。IRS/US连接处位于IRS区域第二个EcoRI位点下游950 bp处，而US/TRS连接处位于TRS区域第一个EcoRI限制性酶切位点上游950 bp处。对其中一个PCR产物进行BamHI限制性酶切图谱分析，在MDV基因组的US/TRS连接处上游鉴定出两个新的DNA亚片段，即BamHI-U2和-P4。对BamHI-D片段进行测序，发现了一个编码155个氨基酸的新开放阅读框（ORF）。TRL/UL连接处位于这个ORF中。该蛋白的N端65个氨基酸与先前报道的位于UL/IRL区域的pp38的N端区域同源。计算机辅助分析表明，两者都是跨膜蛋白，并且它们共享一个抗原结构域。
