Artavanis-Tsakonas S, Schedl P, Tschudi C, Pirrotta V, Steward R, Gehring W J
Cell. 1977 Dec;12(4):1057-67. doi: 10.1016/0092-8674(77)90169-6.
We have cloned embryonic Drosophila DNA using the poly (dA-DT) connector method (Lobban and Kaiser, 1973) and the ampicillin-resistant plasmid pSF2124 (So, Gill and Falkow, 1975) as a cloning vehicle. Two clones, containing hybrid plasmids with sequences complementary to a 5S RNA probe isolated from Drosophila tissue culture cells, were identified by the Grunstein and Hogness (1975) colony hybridization procedure. One hybrid plasmid has a Drosophila insert which is comprised solely of tandem repeats of the 5S gene plus spacer sequences. The other plasmid contains an insert which has about 20 tandem 5S repeat units plus an additional 4 kilobases of adjacent sequences. The size of the 5S repeat unit was determined by gel electrophoresis and was found to be approximately 375 base pairs. We present a restriction map of both plasmids, and a detailed map of of the5S repeat unit. The 5S repat unit shows slight length and sequence heterogeneity. We present evidence suggesting that the 5S genes in Drosophila melanogaster may be arranged in a single continuous cluster.
我们使用聚（dA-DT）连接子法（洛班和凯泽，1973年）以及氨苄青霉素抗性质粒pSF2124（索、吉尔和法尔科夫，1975年）作为克隆载体，克隆了果蝇胚胎DNA。通过格伦斯坦和霍格内斯（1975年）的菌落杂交程序，鉴定出两个克隆，它们包含与从果蝇组织培养细胞中分离出的5S RNA探针具有互补序列的杂交质粒。一个杂交质粒有一个果蝇插入片段，该片段仅由5S基因的串联重复序列加上间隔序列组成。另一个质粒包含一个插入片段，它有大约20个串联的5S重复单元以及另外4千碱基的相邻序列。5S重复单元的大小通过凝胶电泳确定，发现约为375个碱基对。我们给出了两个质粒的限制性图谱以及5S重复单元的详细图谱。5S重复单元显示出轻微的长度和序列异质性。我们提供的证据表明，黑腹果蝇中的5S基因可能排列成一个单一的连续簇。
