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Abstract:

In order to obtain the cDNA of ClpP gene and analyze the relationships between ClpP gene and virescent mutation of cucumber, the total RNA was extracted from virescent mutational buds of cucumber by TRNzol method. The total RNA was used as template to synthesis the first chain of cDNA by reverse transcription approach. One pair of primers was designed and synthesized to amplify an aim fragment according to relatively ClpP gene sequence. The aim fragment was inserted to a plant expression vector after sequenced. The complete cDNA sequence was gain and submited to NCBI database, which length is 495 bp and encode 158 aa. The predicted protein is about41.00356 Kda, and its isoelectric point is about 5.22. It without signal peptide but included a S14_ClpP_2 conservative structure domain. The amino acids sequence of Clp protease and the amino acids sequence of Clp protease of buttercup family have a high homology. A plant expression vector pBI121-ClpP which has a CaMV 35S prompte was also successfully constructed in this essay. The results demonstrated the ClpP gene exsited in cucumber, and ClpP gene was cloned in cucumber firstly and its whole sequence was got.