Abstract
MicroRNAs (miRNAs) are small RNAs widely present in animals and plants and involved in post-transcriptional regulation of gene transcripts. In this study we identified and validated 58 miRNAs from an EST dataset of Spodoptera litura based on the computational and experimental analysis of sequence conservation and secondary structure of miRNA by comparing the miRNA sequences in the miRbase. RT-PCR was conducted to examine the expression of these miRNAs and stem-loop RT-PCR assay was performed to examine expression of 11 mature miRNAs (out of the 58 putative miRNA) that showed significant changes in different tissues and stages of the insect development. One hundred twenty eight possible target genes against the 11 miRNAs were predicted by using computational methods. Binding of one miRNA (sli-miR-928b) with the three possible target mRNAs was confirmed by Southern blotting, implying its possible function in regulation of the target genes.
Highlights
More than 15000 miRNAs have been identified from different species, such as Bombyx mori, Caenorhabditis elegans, Arabidopsis thaliana and Homo sapiens, by either computational or experimental method and deposited in the miRbase
These miRNAs were identified from 1,132 ESTs, which have nohit homologues in the public databases, by sequence comparison with the 403 known miRNAs of insects, including B. mori, A. gambiae, D. pseudoobscura, D. melanogaster and A. mellifera
In stem-loop RT-PCR the primers were designed based on specific miRNA structure, only those miRNAs that have the same nucleotide sequences as the primers can be amplified by RT-PCR
Summary
Since the second miRNA let-7 was reported in 2000 [2], these vital participators in post-transcriptional gene regulation have received more and more attention and many efforts have been made to discover new miRNAs in different species. MiRNA plays important roles in many physiological processes, such as growth, development, metabolism, behavior and apoptosis by mRNA cleavage or translational repression [1,3,4,5,6,7,8,9]. It is found that one miRNA can target mRNA of several genes. MRNAs of one-third of genes are regulated by miRNA as transcriptional or developmental factor; on the other hand, one molecule of mRNA can be bound by several different miRNAs [10]
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
