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Construction of Sly-miR393 Over-Expression Vector and Verification of Its Target Genes in Tomato

Dongbo Lin, Yingwu Yang, Zhengguo Li

Article ID: 1169
Vol 4, Issue 1, 2020, Article identifier:

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To understand the function of Sly-miR393 in tomato, the precursor sequences and potential target genes of Sly-miR393 were identificated from tomato genome database by computational homology search method. The Sly-miR393 gene was amplified from the genomic DNA by PCR and cloned into plant expression vector pLP35s-100. Sly-miR393 guided-cleavage to putative target  transcripts was validated u sing 5RACE RT-PCR. In this study, our results indicated that the precursor sequence of Sly-miR393 contains the complete hairpin  structure. TIR1/AFB auxin receptor genes contain recognition sites with high complementarities to Sly-miR393 sequence. In tomato, Sly-miR393 directs the cleavage of SlTIR1,SlTIR1-like1 and SlAFB mRNA, then auxin receptor homologous was validated to be as target of Sly-miR393. The pLP35s-pre-SlymiR393 vector containing Sly-miR393 gene was successfully constructed, which would provide significant evidence for further study of Sly-miR393 function in auxin signaling pathway in tomato.


Sly-miR393; Target Genes; Auxin Receptor Gene; Construction

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