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A prespore-specific gene of Dictyostelium discoideum encodes the small subunit of ribonucleotide reductase

机译:盘基网柄菌的芽孢前特异性基因编码核糖核苷酸还原酶的小亚基

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摘要

We have isolated the gene, rnrB, that encodes the ribonucleotide reductase small subunit of Dictyostelium discoideum. The deduced amino acid sequence of rnrB exhibits about 60% sequence identity with its homologues in other eukaryotes. As demonstrated by RNA blot analysis the rnrB transcript is detected in growing cells and decreases dramatically at the onset of development. The rnrB transcript reappears after the cells have formed multicellular aggregates. Xo further examine the pattern of expression, we have fused the rnrB promoter and part of its coding sequence to lacTL. The; transgenic strain bearing such a reporter construct expresses the fusion gene with a biphasic profile, which is indistinguishable from that of the endogenous rnrB. The multicellular aggregates of Dictyostelium are differentiated along the anterior-posterior axis. Cells in the anterior give rise to the stalk of the fruiting body while cells in the posterior are precursors of spores.; Results from histochemical staining show that (3-galactosidase activity is detected exclusively in the posterior two-thirds of I the aggregates. These data suggest that rnrB is expressed in prespore cells during postaggregative development and in vegetative cells.
机译:我们已经分离了编码盘基网柄菌的核糖核苷酸还原酶小亚基的rnrB基因。推导的rnrB氨基酸序列与其在其他真核生物中的同源物具有约60%的序列同一性。正如RNA印迹分析所证实的那样,在生长中的细胞中检测到rnrB转录物,并在发育开始时急剧下降。细胞形成多细胞聚集体后,rnrB转录物重新出现。 Xo进一步检查了表达模式,我们将rnrB启动子及其部分编码序列与lacTL融合。的;带有这种报告基因构建体的转基因菌株表达的融合基因具有双相特征,与内源性rnrB的特征没有区别。 Dictyostelium的多细胞聚集体沿前后轴分化。前部的细胞产生子实体的茎,而后部的细胞则是孢子的前体。组织化学染色的结果表明(3-半乳糖苷酶活性仅在聚集体的后三分之二处检测到。这些数据表明rnrB在聚集后发育期间的孢子前细胞和营养细胞中表达。

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