Abstract: Objective: To construct EGFP-Rab7 fusion expression vector, and to investigate its effect on melanin synthesis in A375 cells. Methods: Total RNA was extracted from peripheral lymphocytes for reverse transcription of cDNA. Polychain reaction (PCR) was performed with specific primers to amplify Rab7 fragment. PCR products were digested and inserted into EGFP-C1 vector. The recombinant plasmids EGFP-Rab7 were transformed into E. coli DH5a. Positive clones were screened and identified by DNA sequencing. The recombinant plasmid was transfected into A375 cells and the expression level of melanogenic enzymes and melanin content was detected. Results: EGFP-Rab7 expression vectors were verified by DNA sequencing. The expression level of tyrosinase and tyrosinase-related protein 1 and melanin content in A375 cells transfected by EGFP-Rab7 was higher than that in empty transfer plasmid group. Conclusion: EGFP-Rab7 fusion expression vector is successfully constructed which can increase the level of melanin and and melanin synthetase. 

Key words: Rab7, fusion expression vector, subcellular localization, melanin metabolism