菌株WYT降解还原蓝4的关键基因

以印染厂活性污泥中筛选出的WYT菌株为研究对象，探索其对还原蓝4（VB4）染料降解脱色的关键基因.采用全基因组测序分析方法和同源重组法，确定染料代谢的可能关键基因.对该基因进行敲除，构建载体进行基因回补，设计表型验证实验验证基因的脱色效果.结果表明，该可能关键基因属于染料过氧化物酶基因中的B型，命名为DyP.敲除后的菌株对VB4没有脱色效果，基因敲除成功.WYT菌株与敲除载体发生双交换，获得回补株.在降解实验中，回补株对VB4的脱色率为96.04%，野生株的脱色率为96.95%，回补株恢复了对VB4的降解脱色能力，而敲除株几乎失去对VB4的降解能力，DyP基因是VB4降解脱色的关键基因.

The key genes involved in degradtion of vat blue 4 in bacterial strain WYT

A dye degrading bacteria named WYT was isolated from printing and dyeing activated sludge, and the key genes involved in the degradation and decolorization of VB4dye were investigated. The whole genome sequencing and homologous recombination were used to identify the possible key genes of dye metabolism. The gene was knocked out, meanwhile, the vector was constructed for gene complement, and a phenotypic verification experiment was designed to verify the decolorization effect of the gene. The results showed that the possible key gene, named DyP, belonged to type B of DyP eroxidase gene.The DyP knockout strain had no decolorization effect on VB4. The decolorization rate of VB4 was 96.04% in the complementary strain and 96.95% in the wild type, indicating the decolorization rate of VB4 in the complementary strain was restored, DyP gene was the key gene of VB4 degradation and decolorization.