In this study, Streptomyces avermitilis Bjbm0006 which produces four avermectin B components was used as an original test strain. A replacement plasmid containing a gene cluster bkdAB (branched-chain alpha-keto acid dehydrogenase gene) involved in the biosynthesis of avermectin B in S. avermitilis Bjbm0006 was constructed by means of PCR technique and then named as pHJ5821 (pHZ1358::bkdAB&erm). A recombinant strain Bjbm5821 was obtained after the gene cluster was interrupted by double crossover. This strain was tested in laboratory conditions and analysed by PCR using the total DNA as template. The HPLC analysis showed that the strain Bjbm5821 synthesized the same 'a' components Bla and B2a as the original strain did. However, It lost the ability for the production of 'b'components for example B1b and B2b. A novel compound was detected in fermentation products. The results of present study suggests that the production of gene cluster bkdAB may play a main role similar to alpha-ketoisovaleric acid dehydrogenase in the pathway of avermectin synthesis.