Small-molecule ribonucleic acid (RNA) Osa-miR393 for improving rice tillering and application
A rnaosa-mir393, small molecule technology, applied in the field of plant genetic engineering, can solve problems such as unseen miR393
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[0049] Example 1: Cloning of small RNA Osa-miR393
[0050] Via website http: / / www.mirbase.org Find the sequence of Osa-miR393 and enter this microRNA sequence into the microRNA design application website http: / / wmd3.weigelworld.org / cgi-bin / webapp.cgi? page=Home; project=stdwmd Four primers are automatically synthesized and amplified by multiple PCR to obtain a PCR product fragment expressing the stem-loop structure of Osa-miR393.
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[0051] Example 2: Verification and application of Osa-miR393 function
[0052] 1. Construction of genetic transformation vector
[0053] The vector used for overexpression is Ami393 constructed by our laboratory. Ami393 is a commonly used plant genetic transformation vector pCAMBIA1301 (Sun et al. Xa26, a gene conferring resistance to Xanthomonas oryzae pv.oryzaein rice, encoding a LRR receptor kinase-like protein. Plant Journal. 2004, 37: 517-527). ) Modified on the basis of the Agrobacterium-mediated genetic transformation vector carrying the corn 35S promoter with constitutive and overexpression characteristics ( figure 2 ). The cloned Osa-miR393 fragment was ligated into pGEMT-easy vector, digested with AflII and BglII into the binary expression vector pCambia1301, and the Gus expression gene inside the vector was replaced. The final vector constructed was named Ami393.
[0054] 2. Genetic transformation
[0055] A genetic transformation method mediated by Agrobacterium EHA105...
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