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 photo-protein 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.

Highlights

  • Ribosomal RNA genes are present in multiple copies per genome in all the eukaryotes studied

  • The 5S Ribosomal RNA (rRNA) gene and their associated spacer regions were amplified by using consensus primers complementary to and based on the sequence of 3' [M27 5'-TTTAGTGCTGGTATGATCGC-3'] and 5' [M28 5'TGGGAAGTCCTCGTGTTGCA-3'] ends of 5S rRNA gene coding regions (5S rDNA) from plants as described (Singh and Singh, 2001)

  • The 5S rRNA genes are arranged as tandem repeats in several plant species (Szymanski et al, 2000)

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Summary

Introduction

Ribosomal RNA (rRNA) genes are present in multiple copies per genome in all the eukaryotes studied. The objective of this study, was to clone 5S ribosomal RNA gene(s) from Carica species and to understand their organization in the genome, in order to provide tools to study role of 5S rRNA genes and associated spacers. This will be useful in characterization of huge Carica germplasm as well as in understanding adaptation of these species to temperatures

Plant Material and DNA Isolation
Polymerase Chain Reaction and Cloning of the PCR Products
Sequencing and Sequence Analysis
Gene Cloning Approach
Conclusion
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