Using gene recombinant techniques, two chimeric operons containing rhaSR promoter, RhaR gene, and reporter gene gst (glutathione S-transferase )were constructed, and each was inserted into the E.coli expression vector pALEX to form pALEX-PR1 and pALEX-PR2.The pALEX-PR2 contained a native SD sequence in the upstream region of rhaR, while the pALEX-PR1 contained an enhanced SD sequence. Two plasmids were then transformed into E.coli BL21 (DE3). The reporter gene gst within both chimeric operons expressed 4 to 5 folds higher with L-rhamnose induction than without the induction. Under the induction of L-rhamnose, the GST expression of the pALEX-PR1 was 3.14 folds than the pALEX-PR2.Our results suggested that the expression of gst was positively regulated by the induction of L-rhamnose and the RhaR expressed from the same chimeric operon. Furthermore, SDS-PAGE results showed that GST accounted for about 5.41%(W/W) of the total soluble proteins of the E. coli culture. An average of 3.0 mg purified GST was obtained from 1 L culture. The results of enzyme activity analysis showed that the GST expressed by reporter gene gst of our chimeric operons kept the correct configuration and high activity.