Cloning and Sequence Analysis of 5S Ribosomal RNA Gene(s) and Associated Intergenic Spacer Regions in Carica Species

Mahipal Singh, P. Yadav, A. Yadav
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Abstract

The 5S ribosomal RNA gene(s) and their associated intergenic spacer regions were amplified from Carica papaya and Carica quercifolia by polymerase chain reaction. Both Carica species exhibited differently sized amplification products. Sequence analysis of these PCR products revealed that the 5S rRNA genes are arranged as tandem repeats in these regions. Sequence data revealed that the 5S rRNA gene from Carica quercifolia was 119 bp in length. Sequence variation was observed in various 5S rRNA gene copies cloned from Carica quercifolia. Only truncated 5S rRNA gene but with its full spacer region was recovered from Carica papaya. Interestingly, intergenic spacer sequence cloned from Carica papaya contained two specific domains, a 30bp “CT” rich domain exhibiting 95-100% homology to several human chromosomes and a domain matching with mitrocomin precursor, a photoprotein from Mitrocoma cellularia. The role of 5S rRNA gene and their spacer regions in discerning the germplasm and in adaptation of the species is discussed.
加勒比种5S核糖体RNA基因及相关基因间隔区的克隆与序列分析
采用聚合酶链反应从番木瓜和槲皮中扩增出5S核糖体RNA基因及其相关基因间隔区。两种Carica扩增产物大小不同。PCR产物序列分析表明,5S rRNA基因在这些区域呈串联重复序列排列。序列数据显示,槲皮5S rRNA基因长度为119 bp。从槲皮中克隆的多个5S rRNA基因拷贝序列存在差异。在番木瓜中只发现了截断的5S rRNA基因,但有完整的间隔区。有趣的是,从番木瓜中克隆的基因间间隔序列包含两个特定的结构域,一个是30bp的“CT”丰富结构域,与几种人类染色体具有95-100%的同源性,另一个是与mitrocomin前体匹配的结构域,mitrocomin前体是一种来自Mitrocoma cellularia的光蛋白。讨论了5S rRNA基因及其间隔区在种质识别和物种适应中的作用。
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