Abstract:

To investigate the expected functions of the SAD from Camellia oleifera seeds, a pair of primer integrated with two restriction sites was designed according to the reported coding sequence (CDS) of the SAD, thereafter the segment of CDS, which contained double digestion sites, was obtained by RT-PCR and TA cloning method. After confirmation of the CDS by PCR and sequencing respectively, the PCR products of the CDS was double digested and ligated to pET30a which was aslo double digested using the same restriction enzymes, followed by transformation of recombinant pET-CoSAD into BL21 (DE3). The length of colony PCR products was the same as the expected, and the distribution of double digested products via electrophoresis was also in accordance with experiential distribution. In summary, a recombinant pET-CoSAD which could be used to analyze the function of CoSAD in pET system had been constructed successfully.