Powell A L, King W, Kieff E
J Virol. 1979 Jan;29(1):261-74. doi: 10.1128/JVI.29.1.261-274.1979.
Namalwa and Raji cells, originally obtained from a Burkitt tumor biopsy, grow as continuous cell lines in vitro and contain the Epstein-Barr virus (EBV)-related nuclear antigen EBNA (B. M. Reedman and G. Klein, Int. J. Cancer 11:499-520, 1973) and RNA homologous to at least 17 and 30% of the EBV genome, respectively (S. D. Hayward and E. Kieff, J. Virol. 18:518-525, 1976; T. Orellana and E. Kieff, J. Virol. 22:321-330, 1977). The polyribosomal and polyadenylated [poly(A)+] RNA fractions of Namalwa and Raji cells are enriched for a class of viral RNA homologous to 5 to 7% of EBV DNA (Hayward and Kieff, J. Virol. 18:518-525, 1976; Orellana and Kieff, J. Virol. 22:321-330, 1977). The objective of the experiments described in this communication was to determine the location within the map of the EBV genome (D. Given and E. Kieff, J. Virol. 28:524-542, 1978) of the DNA which encodes the viral RNA in the poly(A)+ and non-polyadenylated [poly(A)-] RNA fractions of Namalwa cells. Hybridization of labeled DNA homologous to Namalwa poly(A)+ or poly(A)- RNA to blots containing EcoRI, Hsu I, or Hsu I/EcoRI double-cut fragments of EBV (B95-8) or (W91) DNA indicated that these RNAs are encoded by DNA contained primarily in the Hsu I A/EcoRI A and Hsu I B/EcoRI A fragments and, to a lesser extent, in other fragments of the EBV genome. Hybridizations of Namalwa poly(A)+ and poly(A)- RNA in solution to denatured labeled EcoRI A or B fragments, Hsu I A, B, or D fragments, and Hsu I A/EcoRI A or Bam I S fragments and of Raji polyribosomal poly(A)+ RNA to the EcoRI A fragment indicated that (i) Namalwa poly(A)+ RNA is encoded primarily by 6 x 10(5) daltons of a 2 x 10(6)-dalton segment of DNA, Bam I S, which is tandemly reiterated, approximately 10 times, in the Hsu I A/EcoRI A fragment and is encoded to a lesser extent by DNA in the Hsu I B, EcoRI B, and Hsu I D fragments. Raji polyribosomal poly(A)+ RNA is encoded by a similar fraction of the EcoRI A fragment as that which encodes Namalwa poly(A)+ RNA. (ii) The fraction of the Bam I S fragment homologous to Namalwa poly(A)- RNA is similar to the fraction homologous to Namalwa poly(A)+ RNA. However, Namalwa poly(A)- RNA is homologous to a larger fraction of the DNA in the Hsu I B, Hsu I D, and EcoRI B fragments.
Namalwa细胞和Raji细胞最初取自伯基特淋巴瘤活检组织，在体外可作为连续细胞系生长，并含有与爱泼斯坦-巴尔病毒（EBV）相关的核抗原EBNA（B.M.Reedman和G.Klein，《国际癌症杂志》11:499 - 520，1973年）以及分别与EBV基因组至少17%和30%同源的RNA（S.D.Hayward和E.Kieff，《病毒学杂志》18:518 - 525，1976年；T.Orellana和E.Kieff，《病毒学杂志》22:321 - 330，1977年）。Namalwa细胞和Raji细胞的多核糖体和聚腺苷酸化[poly(A)+]RNA组分富含一类与EBV DNA 5%至7%同源的病毒RNA（Hayward和Kieff，《病毒学杂志》18:518 - 525，1976年；Orellana和Kieff，《病毒学杂志》22:321 - 330，1977年）。本论文所述实验的目的是确定在EBV基因组图谱（D.Given和E.Kieff，《病毒学杂志》28:524 - 542，1978年）中，编码Namalwa细胞poly(A)+和非聚腺苷酸化[poly(A)-]RNA组分中病毒RNA的DNA的位置。与Namalwa细胞poly(A)+或poly(A)- RNA同源的标记DNA与包含EBV（B95 - 8）或（W91）DNA的EcoRI、Hsu I或Hsu I/EcoRI双酶切片段的印迹杂交表明，这些RNA主要由包含在Hsu I A/EcoRI A和Hsu I B/EcoRI A片段中的DNA编码，在较小程度上由EBV基因组的其他片段编码。Namalwa细胞的poly(A)+和poly(A)- RNA在溶液中与变性的标记EcoRI A或B片段、Hsu I A、B或D片段以及Hsu I A/EcoRI A或Bam I S片段杂交，Raji细胞的多核糖体poly(A)+ RNA与EcoRI A片段杂交表明：（i）Namalwa细胞的poly(A)+ RNA主要由2×10⁶道尔顿片段中6×10⁵道尔顿的DNA编码，该片段为Bam I S，在Hsu I A/EcoRI A片段中串联重复约10次，在较小程度上由Hsu I B、EcoRI B和Hsu I D片段中的DNA编码。Raji细胞的多核糖体poly(A)+ RNA由与编码Namalwa细胞poly(A)+ RNA的EcoRI A片段相似的部分编码。（ii）与Namalwa细胞poly(A)- RNA同源的Bam I S片段部分与与Namalwa细胞poly(A)+ RNA同源的部分相似。然而，Namalwa细胞的poly(A)- RNA与Hsu I B、Hsu I D和EcoRI B片段中更大比例的DNA同源。
