Rekosh D, Lindenbaum J, Brewster J, Mertz L M, Hurwitz J, Prestine L
Proc Natl Acad Sci U S A. 1985 Apr;82(8):2354-8. doi: 10.1073/pnas.82.8.2354.
The bulk of an open reading frame extending from map coordinates 23.3 to 14.2 in region E2b of the adenoviral genome has been cloned and expressed from a chimeric plasmid in Escherichia coli. The cloning strategy used created a fusion protein of 124,000 daltons, which contained greater than 98% adenovirus-encoded sequences. Antiserum raised against this protein reacted with the authentic 140,000-dalton adenovirus DNA polymerase. Another serum raised against a synthetic hexapeptide whose sequence corresponded to the predicted carboxyl terminus of adenovirus-encoded DNA polymerase also reacted with the fusion protein and authentic adenovirus DNA polymerase. These results demonstrate that the cloned region of DNA encodes the adenovirus DNA polymerase.
从腺病毒基因组E2b区图谱坐标23.3至14.2延伸的大部分开放阅读框已从嵌合质粒中克隆出来,并在大肠杆菌中表达。所采用的克隆策略产生了一种124,000道尔顿的融合蛋白,其中包含超过98%的腺病毒编码序列。针对该蛋白产生的抗血清与真实的140,000道尔顿腺病毒DNA聚合酶发生反应。另一种针对合成六肽产生的血清,其序列与腺病毒编码的DNA聚合酶预测的羧基末端相对应,也与融合蛋白和真实的腺病毒DNA聚合酶发生反应。这些结果表明,DNA的克隆区域编码腺病毒DNA聚合酶。