McClements W, Hanafusa H, Tilghman S, Skalka A
Proc Natl Acad Sci U S A. 1979 May;76(5):2165-9. doi: 10.1073/pnas.76.5.2165.
DNA from a variety of uninfected chicken cell types has been analyzed by using restriction endonuclease digestion and RPC-5 ion-exchange chromatography followed by agarose gel electrophoresis. Endogenous retrovirus sequences were detected by using a 32P-labeled avian leukosis viral RNA probe. One simple pattern was identified in an individual containing unexpressed endogenous proviral genes (gs-chf-phenotype for group-specific antigens and chicken helper factor) that was common to all individuals studied. A tentative restriction has been derived for this and one other gs-chf-endogenous provirus. Other gs-chf-individuals and individuals with other phenotypes (e.g., gs+ chf+ and gsl chlfhE) showed more complicated patterns that often included additional bands and thus probably additional proviruses. RNA from an avian sarcoma virus was used to detect cellular sequences (sarc) homologous to the viral transforming gene (src). Results have revealed that a single restriction endonuclease EcoRI fragment of 13 x 10(6) daltons contains the majority of these sequences and confirm that they are not adjacent to the endogenous provirus.
利用限制性内切酶消化、RPC - 5离子交换色谱法,随后进行琼脂糖凝胶电泳,对来自多种未感染鸡细胞类型的DNA进行了分析。使用32P标记的禽白血病病毒RNA探针检测内源性逆转录病毒序列。在一个含有未表达内源性前病毒基因(群特异性抗原和鸡辅助因子的gs - chf - 表型)的个体中鉴定出一种简单模式,该模式在所有研究个体中都很常见。已初步确定了这种和另一种gs - chf - 内源性前病毒的限制性图谱。其他gs - chf个体和具有其他表型(例如,gs + chf + 和gsl chlfhE)的个体显示出更复杂的模式,通常包括额外的条带,因此可能含有额外的前病毒。使用禽肉瘤病毒的RNA来检测与病毒转化基因(src)同源的细胞序列(sarc)。结果表明,一个13×10(6)道尔顿的单一限制性内切酶EcoRI片段包含了这些序列的大部分,并证实它们不与内源性前病毒相邻。