Abstract: To reveal the pathway of miRNA regulating tomato different growth and development stage, especially in fruit ripening, the research obtained tomato (Solanum lycopersicum) miR172 gene by the polymerse chain reaction technique (PCR), then we used the restriction enzymes to digest especially gene segments and pCAMBIA1300-221 vector, and connected the gene to the plasmid pCAMBIA1300-221, then transformed into Agrobacterium by Freeze-thaw method when the recombinant plasmid was accurate. The recombinant plasmid was transformed again into Escherichia coli in order to obtain the exact plasmid by PCR and double enzyme digestion in the research, and the sequence result was homology to NCBI database up to 100%. The results indicated that tomato expression vector pCAMBIA1300-221-miR172 was constructed successfully and then we cultivated and identificate the transgenic plants. It would provide base for genetic research in fruit ripening and aging through miR172 transgenic tomatoes.