Persimmon ACC synthase gene cloning and its plant expression vector construction

An amino-cyclopropane-1-carboxylie acid synthase(ACS) gene was amplified by Reverse Transcription Poly-merse Chain Reaction (RT-PCR) from ripening persimmon fruit. A 1 178 bp PCR product (Fuyu-ACS) was cloned. Sequence analysis showed that Fuyu-ACS nucleotide sequence was 99% identity with DK-ACS1 in GenBank. Two pairs of primers containing restriction enzyme site were designed and were used to amplify sequenced plasmid. Two PCR products were digested by the corresponding restricted enzymes respectively, and inserted between the CaMv 35 S promoter and NOS terminator of expression vector pBI121.