To investigate the role of  lincomycin transporter LmrC in lincomycin biosynthesis, lmrC was deleted in a lincomycin high-yielding strain LC-G by homologous recombination. HPLC analysis and resistance assay indicated that lincomycin production and self-resistance in XJJ7, and the lmrC mutant were decreased by 55% and 50%, respectively. Quantitative real-time polymerase chain reaction(PCR) analysis showed that the transcriptional levels of structure genes lmbA and lmbR were drastically reduced, whereas the expression of a regulatory gene lmbU did not change in XJJ7. Moreover, overexpression of lmrC in LCG could not increase the lincomycin yield and the expression of lmbA, lmbR and lmbU as well. However, the resistance to lincomycin was improved in the lmrCoverexpressed strain. Therefore, lmrC was proved to be involved in lincomycin biosynthesis by affecting the transcription of structure genes and confer lincomycin resistance to Streptomyces lincolnensis.