Cloning and Expression Vector Construction of CbP5CS1 Gene from Cinna-momun bodinieri
In order to study the structure and function of the CbP5CS1 protein sequence,based on the transcrip-tome data of Cinnamomun bodinieri,the CDS gene was cloned by PCR and its expression vector was constructed.The results showed that the sequence had a complete ORF frame of 2 163 bp encoding 720 amino acids,which was 98.47％consistent with the sequence of RWR86140.1 gene from Cinnamomun micranthum,and the GenBank entry number was MW813958;bioinformatics analysis of CbP5CS1 protein showed that the molecular formula of CbP5CS1 protein was C3435H5609N963O1050S22,the relative molecular weight was 77.91 kD,and the theoretical isoelec-tric point was 6.16.CbP5CS1 protein was fat soluble and hydrophilic without obvious disorder characteristics,and it had 95 phosphorylation amino acid sites;CbP5CS1 protein has the PLN02418 family domain and has the func-tion of △1-pyrrolin-5-carboxylate synthetase,which showed high conserved in Cinnamomum species;CbP5CS1 pro-tein was predicted to be cytoplasmic and transmembrane protein by subcellular localization,but there was no signal peptide;after the recombinant plasmid was digested with XbaI enzyme,two bands were obtained,which were the same size as expected 2 226 and 11 516 bp,indicating that the target gene had been successfully inserted and at-tached to the plant expression vector pCAMBIA1301.
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