Cloning and Sequencing of NorB Gene of Nitrobacterium / 环境与健康杂志

ABSTRACT

Objective To amplify and sequence the distinctive norB gene of Nitrobacterium. Methods The norB gene was amplified by PCR and was cloned into T-vector and then transformed into E.coli JM109. After white-and-blue selection, positive colonies were sequenced with T7 sequencing primers by Takara Company. Spliced with Vector NTI Suite software, the homologous analysis of sequences were conducted in Genbank through Internet. Results The results showed that all of norB genes obtained from the nitrite-oxidizing bacteria were homologous with the norB gene of Nitrobacterium hamburgensis in Genbank. The homology ranged from 96% to 98%, and there were no frameshift mutations, which guaranteed the correct ORF. Conclusion The obtained genes are norB genes indeed.