ABSTRACT
Objective: To identify and highly express the L-Methionine ?-lyase gene from Trichomonas vaginalis(TVMGL).Methods: The TVMGL gene was cloned into pGEX 4T-2,the recombinant protein expressed in E.coli DH5?and confirmed by SDS-PAGE.Results: Denaturing SDS-PAGE analysis confirmed the predicted size of the fusion protein(GST-TVMGL,68 000) and showed high purity after subsequent affinity chromatography on Glutathione sepharose 4B.The target proteins amounted to 34% of the total bacteria proteins.Conclusion: The L-Methionine ?-lyase gene was highly expressed in E.coli.
ABSTRACT
Objective:To identify,clone,sequence the L-Methionine ?-lyase gene from Trichomonas vaginalis(TVMGL1).Methods:Total RNA was prepared from primitive protozoon trichomonas vaginalis,cDNA fragment encoding Methionine ?-lyase gene was amplified by RT-PCR using specific primers,and then was cloned into pGEM-T vector.Inserted Methionine ?-lyase gene was sequenced by ABI 3730 DNA Sequencer.Results:Analysis indicates that the DNA fragment is 1 191 base pairs in length and has high nucleotide homology with that reported previously.Conclusion:L-Methionine ?-lyase gene was successfully cloned.