Study on self-splicing of split intein of the DNA polymerase DnaE in cyanobacteria

【Objective】The double recombinant expression plasmid was cloned to determine the self-splicing activity of split inteins located in split DnaE of cyanobacteria Anabaena sp.PCC 7120.【Method】The corresponding antiserum against the two split DnaE precursor proteins were collected by injecting rabbits with purified proteins after over-expression from reconstructing recombinant expressing plasmid in Escherichia coli.In heterogeous E.coli,the antibodies were available to test the self-splicing of split DnaEs.【Result】Results showed that both precursor proteins and mature proteins were detected in E.coli and antibodies could only react with their corresponding proteins.【Conclusion】The results suggested that trans-splicing existed between the split DnaEs which contain the split inteins.The antibodies were specific without odvious background disturbance and had high effect with applied concentration low to 1/1 000,so it could be used in related research.