Tobin S L, Zulauf E, Sánchez F, Craig E A, McCarthy B J
Cell. 1980 Jan;19(1):121-31. doi: 10.1016/0092-8674(80)90393-1.
In this paper we describe the isolation and characterization of a 7.2 kb D. melanogaster chromosomal DNA fragment (K1) which contains nucleotide sequences complementary to D. melanogaster actin mRNA. Plasmid K1 was identified using a Dictyostelium actin cDNA plasmid, B1, as a probe. D. melanogaster mRNA selected by hybridization with immobilized K1 DNA was translated in vitro to yield products which co-migrate with the D. melanogaster actins I, II and III in two-dimensional gel electrophoresis and bind to DNAase I agarose. A physical map localizing restriction endonuclease cleavage sites in the K1 DNA fragment and the direction of transcription is presented. The position of the coding region has been localized by hybridization with labeled B1 DNA and with labeled poly(A)-containing D. melanogaster RNA. On the basis of hybridization of labeled subfragments of plasmid K1 to restriction endonuclease-cleaved D. melanogaster embryo DNA, we conclude that the nucleotide sequence of the presumptive coding region is responsible for labeling of a pattern of multiple restriction fragments from embryo DNA. The chromosomal locus from which DNA fragment K1 is derived has been localized by in situ hybridization to two closely linked bands in the region 88F. Related DNA sequences corresponding to putative actin genes have also been mapped cytologically. These results support the hypothesis that the genes for actin in D. melanogaster are members of a closely related family of coding sequences.
在本文中，我们描述了一个7.2 kb的黑腹果蝇染色体DNA片段（K1）的分离和特性，该片段包含与黑腹果蝇肌动蛋白mRNA互补的核苷酸序列。使用盘基网柄菌肌动蛋白cDNA质粒B1作为探针鉴定出质粒K1。与固定化的K1 DNA杂交选择的黑腹果蝇mRNA在体外进行翻译，产生的产物在二维凝胶电泳中与黑腹果蝇肌动蛋白I、II和III共同迁移，并与DNA酶I琼脂糖结合。给出了定位K1 DNA片段中限制性内切酶切割位点和转录方向的物理图谱。通过与标记的B1 DNA和标记的含poly(A)的黑腹果蝇RNA杂交，确定了编码区的位置。基于质粒K1标记亚片段与限制性内切酶切割的黑腹果蝇胚胎DNA的杂交，我们得出结论，推测编码区的核苷酸序列负责标记胚胎DNA中多个限制性片段的模式。通过原位杂交将DNA片段K1衍生的染色体位点定位到88F区域的两个紧密相连的带。与假定肌动蛋白基因相对应的相关DNA序列也已在细胞学上进行了定位。这些结果支持了这样的假设，即黑腹果蝇中的肌动蛋白基因是一个紧密相关的编码序列家族的成员。
