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The beta-gal coding region was optimized by codon usage analysis and used as a reporter to monitor the stability and subcellular localization of the expressed eCG protein. In CHO cells, the beta-gal coding region was localized in both the nucleus and cytoplasm. The proportion of the eCG protein in the cytoplasm was approximately 7%. In contrast, in CHO-K1 cells, the eCG protein was expressed at a lower level. The proportion of the beta-gal protein was approximately 55% and 65% in the nucleus and cytoplasm, respectively. Western blot analysis revealed that the EGFP-eCG fusion protein was expressed in the medium at a low level in CHO cells and was secreted into the medium. The eCG fusion protein was mainly localized in the medium and subcellular membranes. We speculate that the different cellular distribution of the beta-gal protein may be due to the translation initiation of the two genes in bicistronic mRNA. 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