Nagao R T, Shah D M, Eckenrode V K, Meagher R B
DNA. 1981;1(1):1-9. doi: 10.1089/dna.1.1981.1.1.
We have investigated the actin-related sequences in soybean using heterologous actin DNA probes from Dictyostelium, Drosophila, and yeast. Southern blot analysis of restriction digests of soybean DNA indicates that actin is encoded in a small multigene family. In order to isolate individual members of this gene family, we have constructed a soybean genomic library in the lambda vehicle Charon 4A. A partial characterization of this library shows it to be nearly complete. We have isolated from this library a number of recombinant clones that hybridize to actin-coding sequences from all three heterologous probes. We have identified the fragments containing the actin-related sequences on the physical maps of two of these clones lambda SAc1 and lambda SAc3. These fragments were subcloned in the plasmid vehicle pBR322. Using electron microscope heteroduplex mapping we show that the subclones, pSAc1 and pSAc3, share homology with the entire actin-coding sequence (1.1 kb) of Drosophila and Dictyostelium. Furthermore, pSAc1 and pSAc3 have additional homology of approximately 0.22 kb at the 5' ends of their coding sequences. No homology is detected in the 3' flanking regions of these clones. The actin sequence in pSAc1 contains an interruption of approximately 0.30 kb located 0.39 kb from the 5' end of the actin polypeptide coding region.
我们使用来自盘基网柄菌、果蝇和酵母的异源肌动蛋白DNA探针，研究了大豆中与肌动蛋白相关的序列。对大豆DNA限制性酶切片段的Southern印迹分析表明，肌动蛋白由一个小的多基因家族编码。为了分离该基因家族的各个成员，我们构建了一个以λ载体Charon 4A为载体的大豆基因组文库。对该文库的部分特征分析表明它几乎是完整的。我们从这个文库中分离出了许多重组克隆，它们能与来自所有三种异源探针的肌动蛋白编码序列杂交。我们已经在其中两个克隆λSAc1和λSAc3的物理图谱上鉴定出了含有肌动蛋白相关序列的片段。这些片段被亚克隆到质粒载体pBR322中。使用电子显微镜异源双链图谱分析，我们发现亚克隆pSAc1和pSAc3与果蝇和盘基网柄菌的整个肌动蛋白编码序列（1.1 kb）具有同源性。此外，pSAc1和pSAc3在其编码序列的5'端还有约0.22 kb的额外同源性。在这些克隆的3'侧翼区域未检测到同源性。pSAc1中的肌动蛋白序列在距肌动蛋白多肽编码区5'端0.39 kb处有一个约0.30 kb的中断。
