Ahmed C M, Chanda R S, Stow N D, Zain B S
Gene. 1982 Dec;20(3):339-46. doi: 10.1016/0378-1119(82)90202-5.
The cytoplasmic poly(A)RNA from early stages of infection of HeLa cells by adenovirus 2 (Ad2), was used to synthesize cDNA. The resulting cDNA segments were inserted at the PstI site of the plasmid pBR322 after dG/dC tailing. The clones containing sequences corresponding to early region III transcripts were identified by hybridization against the SmaI-C fragment (76.5 to 91.9 map units) of the Ad2 genome. The clone pE22 which contained a copy of the mRNA coding for the Mr 19 000 glycoprotein was characterized in detail. A comparison with the genomic sequences allowed the identification of the splice between the x and y leaders. The nucleotide sequences CCGGTG and CAGTTT were found at the donor and acceptor sites of the splice junction, respectively. The coding region consisted of a continuous stretch of 159 amino acids with a hydrophobic N-terminus and two possible glycosylation sites. The triplet ATG was encountered twice in phase 3 before the actual site of initiation of translation, which was in phase 2. The 3'-untranslated region was 500 nucleotides long and contained an open, translational reading frame for a Mr 11 000 protein, following a potential initiator ATG. The sequence ATTAAA was observed 17 bp before the poly(A) tail.
用腺病毒2(Ad2)感染HeLa细胞早期阶段的细胞质多聚腺苷酸RNA来合成cDNA。在进行dG/dC加尾后,将得到的cDNA片段插入质粒pBR322的PstI位点。通过与Ad2基因组的SmaI-C片段(76.5至91.9图谱单位)杂交,鉴定出含有与早期区域III转录本相对应序列的克隆。对包含编码19000 Mr糖蛋白的mRNA拷贝的克隆pE22进行了详细表征。与基因组序列的比较使得能够鉴定出x和y前导序列之间的剪接。在剪接连接的供体和受体位点分别发现了核苷酸序列CCGGTG和CAGTTT。编码区由连续的159个氨基酸组成,具有疏水的N末端和两个可能的糖基化位点。三联体ATG在实际翻译起始位点之前的相位3中出现了两次,而实际翻译起始位点处于相位2。3'-非翻译区长度为500个核苷酸,在一个潜在的起始ATG之后,包含一个针对11000 Mr蛋白质的开放阅读框。在多聚腺苷酸尾巴之前17 bp处观察到序列ATTAAA。