Wilson G N, Steggles A W, Nienhuis A W
Proc Natl Acad Sci U S A. 1975 Dec;72(12):4835-9. doi: 10.1073/pnas.72.12.4835.
In order to investigate the symmetry of globin gene transcription, complementary RNA (cRNA) was synthesized using rabbit globin complementary DNA (cDNA) as a template for Escherichia coli DNA-dependent RNA polymerase (RNA nucleotidyltransferase). The cRNA hybridized specifically to its own cDNA template but not to sheep cDNA, rabbit globin mRNA, or poly(dT). Hybridization studies with cRNA demonstrated that RNA sequences transcribed from the DNA strand complementary to the globin gene region (anti-strand) were not present in cellular, total nuclear, or fractionated nuclear RNA from rabbit marrow. Such sequences were detected in RNA transcribed from rabbit marrow chromatin by E. coli or sheep liver RNA polymerases, but amounted to less than 50% of the globin mRNA sequences present in the same transcript. The evidence indicates that globin mRNA transcription is predominantly DNA strand specific.
为了研究珠蛋白基因转录的对称性,以兔珠蛋白互补DNA(cDNA)为模板,利用大肠杆菌DNA依赖性RNA聚合酶(RNA核苷酸转移酶)合成了互补RNA(cRNA)。cRNA与其自身的cDNA模板特异性杂交,但不与绵羊cDNA、兔珠蛋白mRNA或聚(dT)杂交。用cRNA进行的杂交研究表明,从与珠蛋白基因区域互补的DNA链(反义链)转录的RNA序列不存在于兔骨髓的细胞RNA、总核RNA或分级分离的核RNA中。通过大肠杆菌或绵羊肝RNA聚合酶从兔骨髓染色质转录的RNA中检测到了此类序列,但在同一转录本中其含量不到珠蛋白mRNA序列的50%。证据表明珠蛋白mRNA转录主要是DNA链特异性的。