Chambers J A, Cywinski A, Chen P J, Taylor J M
J Virol. 1986 Aug;59(2):354-62. doi: 10.1128/JVI.59.2.354-362.1986.
We detected sequences related to the avian retrovirus Rous sarcoma virus within the genome of the Japanese quail, a species previously considered to be free of endogenous avian leukosis virus elements. Using low-stringency conditions of hybridization, we screened a quail genomic library for clones containing retrovirus-related information. Of five clones so selected, one, lambda Q48, contained sequence information related to the gag, pol, and env genes of Rous sarcoma virus arranged in a contiguous fashion and spanning a distance of approximately 5.8 kilobases. This organization is consistent with the presence of an endogenous retroviral element within the Japanese quail genome. Use of this element as a high-stringency probe on Southern blots of genomic digests of several quail DNA demonstrated hybridization to a series of high-molecular-weight bands. By slot hybridization to quail DNA with a cloned probe, it was deduced that there were approximately 300 copies per diploid cell. In addition, the quail element also hybridized at low stringency to the DNA of the White Leghorn chicken and at high stringency to the DNAs of several species of jungle fowl and both true and ruffed pheasants. Limited nucleotide sequencing analysis of lambda Q48 revealed homologies of 65, 52, and 46% compared with the sequence of Rous sarcoma virus strain Prague C for the endonuclease domain of pol, the pol-env junction, and the 3'-terminal region of env, respectively. Comparisons at the amino acid level were also significant, thus confirming the retrovirus relatedness of the cloned quail element.
我们在日本鹌鹑的基因组中检测到了与禽逆转录病毒劳斯肉瘤病毒相关的序列,日本鹌鹑这一物种此前被认为不含内源性禽白血病病毒元件。利用低严谨度杂交条件,我们在鹌鹑基因组文库中筛选含有逆转录病毒相关信息的克隆。在如此选出的五个克隆中,一个名为λQ48的克隆含有与劳斯肉瘤病毒的gag、pol和env基因相关的序列信息,这些信息以连续的方式排列,跨度约为5.8千碱基。这种组织形式与日本鹌鹑基因组中存在内源性逆转录病毒元件相一致。将该元件用作高严谨度探针,对几种鹌鹑DNA的基因组消化产物进行Southern印迹分析,结果显示与一系列高分子量条带发生杂交。通过用克隆探针与鹌鹑DNA进行狭缝杂交,推断出每个二倍体细胞中约有300个拷贝。此外,鹌鹑元件在低严谨度下也与白来航鸡的DNA杂交,在高严谨度下与几种原鸡以及环颈雉和绿孔雀的DNA杂交。对λQ48进行的有限核苷酸序列分析显示,与劳斯肉瘤病毒布拉格C株相比,pol的核酸内切酶结构域、pol-env连接区和env的3'末端区域的同源性分别为65%、52%和46%。在氨基酸水平上的比较也很显著,从而证实了克隆的鹌鹑元件与逆转录病毒的相关性。