OBJECTIVE: To construct a recombinant plasmid vector of the RV specific fragment for expressing the specific fragment of RV E1 protein.METHODS: RNA of the RV attenuated live vaccine Wistar RA27/3 strain was extracted and reversely transcribed. The specific fragment of the E1 gene was amplified and the PCR products cloned in the vector pGEX-2T after purification. Positive clones were selected and identified by two-enzyme digestion and sequence analysis.RESULTS: A 330 bp target fragment was successfully cloned, and the sequence of the recombinant plasmid was consistent with the original sequence.CONCLUSION: Successful cloning of the RV El specific fragment and the construction of the recombinant plasmid have laid a foundation for further expressing the recombinant protein.
Base Sequence Cloning, Molecular