Construction And Expression Of Prokaryotic Expression Plasmid Encoding Cysticercus Cellulosae Gene CYI

Objective: In order to study the function of cysticercus cellulosae antigen cYI for diagnostic antigen and vaccine preventing cysticercosis, the prokaryotic expression plasmid of cysticercus cellulosae gene cYI was constructed and the expression of fusion protein in E.coli DE3 was observed.Methods:1 The achievement of target gene cYI : Total RNA was extracted from cysticercus cellulosae and RT-PCR was performed using the cYI specific primers and RNA as templates under the following condition of 37℃50min, 94℃5min, then 94℃1min,56℃1min and 72℃1min for 30 cycles and 72°C 10 min after last cycle. The PCR product was digested with Nde I and Xho I, and then identified with 1.0% agarose gel electrophoresis.2 The transformation of recombination plasmid into the E.coli DE3: The resulting RT-PCR product and expression vector pET28b were ligated by T4 DNA ligase after they were digested with Nde I and Xho I, recovered from agarose gel, and purified. The recombination plasmid was then transformed into...