Identification of the gene encoding Marek's disease herpesvirus A antigen
Identification of the gene encoding Marek's disease herpesvirus A antigen
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马立克氏病疱疹病毒 A 抗原编码基因的鉴定
DOI：
10.1128/jvi.61.8.2614-2620.1987
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发表时间：
1987
影响因子：
5.4
通讯作者：
L. F. Velicer
中科院分区：
文献类型：
--
作者：
R. J. Isfort;H. Kung;L. F. Velicer
用Northern印迹(RNA印迹)分析方法，对马立克氏病疱疹病毒A抗原(MDHV-A)前体多肽pr47进行RNA印迹分析，并将其与克隆的MDHV DNA进行杂交筛选、无细胞翻译和免疫沉淀，得到编码MDHV-A前体多肽pr47的基因。杂交选择结果为强阳性的DNA片段是一段2.2kb碱基对(KBP)的PvuII-EcoRI限制性片段，定位于病毒全基因组18.3kbp的MDHV BamHI B片段的中心。这种定位是特异的，因为没有其他较大的BamHI B片段的小限制片段能够杂交选择有意义的MDHV-a mRNA，并且该基因仅定位在病毒总基因组的BamHI B片段中。Northern印迹分析证实MDHV-A基因定位于2.2kbp的片段上，并检测到其mRNA为1.8kb的物种，其大小与编码47kodalton的多肽一致。这是MDHV基因被映射到MDHV病毒基因组的第一份报告。这为利用重组DNA技术研究分泌的病毒特异性糖蛋白基因的性质开辟了道路，该基因可能参与免疫预防、免疫抑制或免疫逃避，即已知或推测与致癌疱疹病毒系统有关的免疫现象。
The gene encoding the glycoprotein Marek's disease herpesvirus A antigen (MDHV-A) precursor polypeptide pr47 was delineated by using Northern blot (RNA blot) analysis and hybrid selection of its mRNA with cloned MDHV DNA, cell-free translation of the mRNA, and immunoprecipitation of the polypeptide. The resulting piece of DNA with strongly positive hybrid selection results was a 2.2-kilobase-pair (kbp) PvuII-EcoRI restriction fragment localized to the center of the 18.3-kbp MDHV BamHI B fragment of the total virus genome. The localization was specific since no other small restriction subfragment of the larger BamHI B fragment was able to hybrid select significant MDHV-A mRNA and the gene mapped only in the BamHI B fragment of the total virus genome. Northern blot analysis confirmed the localization of the MDHV-A gene on the 2.2-kbp fragment and detected its mRNA as a 1.8-kilobase species, a size consistent with encoding a 47-kilodalton polypeptide. This is the first report of an MDHV gene being mapped to the MDHV viral genome. This opens the way for the use of recombinant DNA technology to study the nature of the gene encoding a secreted virus-specific glycoprotein that could possibly be involved in immunoprevention, immunosuppression, or immunoevasion, immune phenomena known or speculated to be involved in this oncogenic herpesvirus system.