Baralle F E
Cell. 1977 Apr;10(4):549-58. doi: 10.1016/0092-8674(77)90088-5.
The 5' noncoding region sequence of rabbit beta-globin mRNA has been determined. This region is 53 nucleotides long, not including the A-U-G initiator sequence or m7Gppp "cap" structure. 32P-labeled DNA complementary to the 5' noncoding region was synthesized using reverse transcriptase with the synthetic deoxyoctanucleotide d(G-C-A-C-C-A-T-T) as a "primer". The cDNA produced was then sequenced using a modification of the gel-sequencing technique previously developed for DNA sequencing (G.G. Brownlee and E.M. Cartwright, manuscript in preparation). The sequence obtained was checked by depurination and nearest-neighbor analysis. The known sequence at the 3' end of rabbit 18S ribosomal RNA cannot base-pair extensively with the 5' noncoding region of beta-globin mRNA; however, it does form 6 base pairs around the initiation codon.
已确定兔β-珠蛋白mRNA的5'非编码区序列。该区域长53个核苷酸,不包括A-U-G起始序列或m7Gppp“帽”结构。以合成的脱氧八聚核苷酸d(G-C-A-C-C-A-T-T)作为“引物”,使用逆转录酶合成了与5'非编码区互补的32P标记DNA。然后,使用先前开发用于DNA测序的凝胶测序技术的改进方法(G.G.布朗利和E.M.卡特赖特,正在准备的手稿)对产生的cDNA进行测序。通过脱嘌呤和邻位分析检查获得的序列。兔18S核糖体RNA 3'端的已知序列不能与β-珠蛋白mRNA的5'非编码区广泛碱基配对;然而,它确实在起始密码子周围形成6个碱基对。