Sprengel J, Schmitz B, Heuss-Neitzel D, Zock C, Doerfler W
Institute of Genetics, University of Cologne, Germany.
J Virol. 1994 Jan;68(1):379-89. doi: 10.1128/JVI.68.1.379-389.1994.
A fresh inoculum of human adenovirus type 12 (Ad12) was obtained from the American Type Culture Collection and passaged once on human embryonic kidney cells, and Ad12 DNA was prepared from the first-passage yield to avoid higher passages which might have generated host-virus DNA recombinants. The 18 PstI fragments of Ad12 DNA were cloned into the pBluescript KS vector, and the entire nucleotide sequence of both strands from all 18 fragments was determined by using successive oligodeoxyribonucleotide primers. Ad12 DNA extends over 34,125 nucleotide pairs, and its molecular weight is calculated to be about 22 x 10(6). The nucleotide sequence of Ad12 DNA was subjected to computer analyses that determined possible open reading of frames on the two strands, the leader sequences, the position of the virus-associated RNA coding region, possible TATA, and polyadenylation signals. The distribution of the Ad12 open reading frames was similar to that in the previously sequenced Ad2 DNA, but there were also distinct differences. Ad12 DNA has an inverted terminal redundancy of 161 nucleotides, compared with 102 nucleotides in Ad2 DNA. There were stretches of sequence identity between Ad2 and Ad12 DNAs at both termini; the overall sequence similarity between the two viral genomes ranged between 59% (polypeptide IX) and 77% (in the E2 region), with high homology also in the sequences for the adenovirus DNA polymerase.
人腺病毒12型(Ad12)的新鲜接种物取自美国典型培养物保藏中心,并在人胚肾细胞上传代一次,从第一代传代产物中制备Ad12 DNA,以避免可能产生宿主-病毒DNA重组体的更高传代。将Ad12 DNA的18个PstI片段克隆到pBluescript KS载体中,并使用连续的寡脱氧核糖核苷酸引物测定所有18个片段两条链的完整核苷酸序列。Ad12 DNA延伸超过34,125个核苷酸对,其分子量计算约为22×10(6)。对Ad12 DNA的核苷酸序列进行计算机分析,确定了两条链上可能的开放阅读框、前导序列、病毒相关RNA编码区的位置、可能的TATA和聚腺苷酸化信号。Ad12开放阅读框的分布与先前测序的Ad2 DNA相似,但也存在明显差异。Ad12 DNA的末端反向重复为161个核苷酸,而Ad2 DNA为102个核苷酸。Ad2和Ad12 DNA在两个末端都有一段序列同一性;两个病毒基因组之间的总体序列相似性在59%(多肽IX)至77%(E2区域)之间,腺病毒DNA聚合酶的序列也具有高度同源性。