携带果蝇tRNA基因的重组DNA质粒的分离与鉴定。

Isolation and characterization of recombinant DNA plasmids carrying Drosophila tRNA genes.

作者信息

Dunn R, Delaney A D, Gillam I C, Hayashi S, Tener G M, Grigliatti T, Misra V, Spurr M G, Taylor D M, Miller R C

出版信息

Gene. 1979 Nov;7(3-4):197-215. doi: 10.1016/0378-1119(79)90046-5.

Abstract

Recombinant plasmids carrying Drosophila melanogaster tRNA genes were constructed by ligation of HindIII-cleaved Drosophila DNA to HindIII cut pBR322 DNA. 90 clones were isolated that contained genes for one or more of eleven tRNAs. 43 of the plasmids were characterized by a number of methods: restriction nuclease digestion; agarose gel electrophoresis; hybridization with individual, purified, 125I-labelled Drosophila tRNA molecules and in situ hybridization to Drosophila chromosomes. The results show that several different tRNA genes have been isolated which code for single, specific isoacceptors. The DNAs from 8 plasmids each hybridize to single sites on Drosophila polytene chromosomes. In addition, the data show examples of two different plasmids hybridizing to different loci coding for the same tRNA; this means that we have isolated representatives of tRNA genes which map at widely separated points on the Drosophila genome.

摘要

通过将经 HindIII 切割的果蝇 DNA 与经 HindIII 切割的 pBR322 DNA 连接，构建了携带黑腹果蝇 tRNA 基因的重组质粒。分离出了 90 个克隆，这些克隆包含 11 种 tRNA 中一种或多种的基因。通过多种方法对 43 个质粒进行了表征：限制性核酸酶消化；琼脂糖凝胶电泳；与单个纯化的 125I 标记的果蝇 tRNA 分子杂交以及与果蝇染色体的原位杂交。结果表明，已分离出几个不同的 tRNA 基因，它们编码单个特定的同功受体。来自 8 个质粒的 DNA 各自与果蝇多线染色体上的单个位点杂交。此外，数据显示了两个不同质粒与编码相同 tRNA 的不同基因座杂交的例子；这意味着我们已经分离出了位于果蝇基因组中广泛分离点的 tRNA 基因的代表。