Wu P, Reed W M, Yoshida S, Sui D, Lee L F
Department of Pathology, Michigan State University, East Lansing, USA.
Acta Virol. 1999 Apr-Jun;43(2-3):152-8.
A 2439 bp open reading frame (ORF) was identified from the DNA sequence of BamHI-F and -K2 fragments of Marek's disease virus of serotype 1 (MDV-1) GA strain, which predicts an 813 amino acid polypeptide. This peptide is homologous to HSV-1 gH, and has typical glycoprotein features. There are nine potential N-linked glycosylation sites within the extracellular domain. A fragment of the gH ORF was cloned into pGEX vector in frame with glutathione S-transferase (GST) to produce a GST-gH fusion protein in Escherichia coli. The GST-gH fusion protein was used to develop gH monoclonal and polyclonal antibodies. Expression of gH was detected in duck embryo fibroblasts (DEFs) infected with MDV-1 GA strain by immunofluorescence assay (IFA) with these antibodies. Virus neutralization and plaque-forming inhibition analyses were conducted with the gH antiserum. There were no neutralization and plaque-forming inhibition activities of gH antiserum. Comparison of the DNA sequence of gH gene between GA and RB1B strains of MDV-1 revealed major difference in the upstream control elements of gH ORF.
从1型马立克氏病病毒（MDV-1）GA株的BamHI-F和-K2片段的DNA序列中鉴定出一个2439 bp的开放阅读框（ORF），该阅读框预测有一个813个氨基酸的多肽。此肽与单纯疱疹病毒1型（HSV-1）gH同源，且具有典型的糖蛋白特征。细胞外区域内有9个潜在的N-糖基化位点。将gH ORF的一个片段与谷胱甘肽S-转移酶（GST）读框内克隆到pGEX载体中，以便在大肠杆菌中产生GST-gH融合蛋白。利用该GST-gH融合蛋白制备gH单克隆抗体和多克隆抗体。用这些抗体通过免疫荧光试验（IFA）在感染MDV-1 GA株的鸭胚成纤维细胞（DEFs）中检测gH的表达。用gH抗血清进行病毒中和及蚀斑形成抑制分析。gH抗血清没有病毒中和及蚀斑形成抑制活性。MDV-1的GA株和RB1B株之间gH基因的DNA序列比较显示，gH ORF的上游控制元件存在主要差异。
