2.3 RNA isolation, cDNA cloning, and sequence analysis
Total RNA from leaves of C. sinensis (SCZ) was isolated using a Fast Pure Plant total RNA Isolation Kit (Vazyme Cat, RC401-01, Nanjing, China) according to the manufacturer’s instructions. The cDNA was synthesized by reverse transcription of the total RNA using Prime Script RT Master Mix (Vazyme, China). The open reading frame sequences were amplified using Phusion ® High-Fidelity DNA Polymerase (New England Biolabs, Ipswich, MA, USA); primers for the clonedCsTPS1/1 -AS gene are shown in Table S1. The PCR products were purified using a Gel Extraction Kit (CWBIO, Jiangsu, China). The resulting target cDNA fragment was ligated into the pGEX-4T1 vector and then subsequently transformed into Trans 1-T1 competent cells.