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鸡肌球蛋白轻链基因的3'非编码区与盘基网柄菌中的一个重复序列家族杂交。

The 3'-noncoding region of the chick myosin light-chain gene hybridizes to a family of repetitive sequences in the slime mold Dictyostelium discoideum.

作者信息

Shaw D R, Khandekar P, Siddiqui M A, Ennis H L

出版信息

Arch Biochem Biophys. 1986 May 1;246(2):829-37. doi: 10.1016/0003-9861(86)90339-5.

Abstract

During studies aimed at isolating myosin-specific genomic clones in Dictyostelium, we probed a lambda genomic library with a chicken myosin light-chain sequence (pML10). Many lambda recombinant Dictyostelium clones hybridized to the pML10 cDNA insert, indicating that this sequence was reiterated in the Dictyostelium genome. It was found that the 3'-noncoding region (pML10-NC) alone was responsible for these results. Dictyostelium DNA contained approximately 65 copies of a sequence(s) similar but not identical to that of pML10-NC. Southern blot analysis showed that pML10-NC hybridized to many Dictyostelium genomic DNA fragments of varying sizes generated by digestion with EcoRI, HindIII, or AluI. In addition, each of the Dictyostelium clones was different in its size, restriction map, and flanking sequences. It seems likely, therefore, that the sequences which hybridized to pML10-NC are scattered throughout the Dictyostelium genome and similar but not identical to each other or to pML10-NC. Thus, probing with pML10-NC has allowed us to select a family of closely related but not identical sequences. These D. discoideum sequences are not found in other slime mold species. No RNA complementary to pML10-NC was found in vegetative cells, 18 h culmination stage, spores, or 1- and 2-h germinating spores. pML10-NC-related sequences were present in two other Dictyostelium species but were absent in the related genus Polysphondylium.

摘要

在旨在分离盘基网柄菌中肌球蛋白特异性基因组克隆的研究过程中,我们用鸡肌球蛋白轻链序列(pML10)探测了一个λ基因组文库。许多λ重组盘基网柄菌克隆与pML10 cDNA插入片段杂交,这表明该序列在盘基网柄菌基因组中是重复的。结果发现,单独的3'-非编码区(pML10-NC)导致了这些结果。盘基网柄菌DNA含有大约65个与pML10-NC序列相似但不完全相同的序列拷贝。Southern印迹分析表明,pML10-NC与用EcoRI、HindIII或AluI消化产生的许多大小不同的盘基网柄菌基因组DNA片段杂交。此外,每个盘基网柄菌克隆在大小、限制性图谱和侧翼序列方面都不同。因此,似乎与pML10-NC杂交的序列分散在盘基网柄菌基因组中,彼此相似但不完全相同,也与pML10-NC不完全相同。因此,用pML10-NC进行探测使我们能够选择一组密切相关但不完全相同的序列。这些盘基网柄菌序列在其他黏菌物种中未发现。在营养细胞、18小时发育阶段、孢子或1小时和2小时萌发的孢子中未发现与pML10-NC互补的RNA。与pML10-NC相关的序列存在于另外两种盘基网柄菌物种中,但在相关的聚孢霉属中不存在。

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