Tsugeki R, Kochieva E Z, Fedoroff N V
Carnegie Institution of Washington, Department of Embryology, Baltimore, MD 21210, USA.
Plant J. 1996 Sep;10(3):479-89. doi: 10.1046/j.1365-313x.1996.10030479.x.
The SSR16 gene of Arabidopsis has been identified as a gene encoding a ribosomal protein S16 homolog through analysis of a transposon insertion mutation. The insertion mutation is lethal, arresting embryonic development at approximately the transition from the globular to the heart stage of embryonic development. Co-segregation of the mutant phenotype with the transposon-borne drug-resistance marker and loss of the inserted transposon concomitant with phenotypic reversion provided evidence that the transposon had caused the mutation. Sequences flanking the insertion site were amplified from DNA of viable heterozygotes by thermal asymmetric interlaced (TAIL) PCR. The amplified fragment flanking the 3' end of the inserted element was sequenced and found to be identical to an Arabidopsis expressed sequence tag (EST). The EST, in turn, contained a coding sequence homologous to the ribosomal protein S16 (RPS16) of bacteria such as Escherichia coli, Bacillus subtilis and Salmonella typhimurium, as well as Neurospora crassa mitochondria and higher plant plastids. Thus the gene identified by the embryo-defective lethal insertion mutation encodes an RPS16 homolog and has been designated the SSR16 gene.
通过对转座子插入突变的分析，拟南芥的SSR16基因已被鉴定为一个编码核糖体蛋白S16同源物的基因。该插入突变是致死性的，在胚胎发育从球状期向心形期转变时使胚胎发育停滞。突变表型与转座子携带的药物抗性标记的共分离以及插入转座子的丢失与表型回复同时出现，提供了转座子导致突变的证据。通过热不对称交错PCR（TAIL-PCR）从存活杂合子的DNA中扩增插入位点侧翼的序列。对插入元件3'端侧翼的扩增片段进行测序，发现其与拟南芥表达序列标签（EST）相同。该EST又包含一个与大肠杆菌、枯草芽孢杆菌和鼠伤寒沙门氏菌等细菌的核糖体蛋白S16（RPS16）以及粗糙脉孢菌线粒体和高等植物质体同源的编码序列。因此，由胚胎缺陷致死插入突变鉴定出的基因编码一个RPS16同源物，并被命名为SSR16基因。
