| 摘要 |
Provided is a method for cloning microbial genes, comprising: using bacterial strains having different target traits as parents, establishing an isolate by hybridization; setting a selection pressure for the isolate to perform automatic filtration, acquiring a post-filtration colony; performing genome high-throughput sequencing on two parents, a pre-filtration colony, and the post-filtration colony; acquiring specific loci of the parents by initial direct assembly and comparison, and comparing between the specific loci of the parents and genomes of the pre-filtration group and of the post-filtration group according to an exact match scheme. A search is performed for a locus that was present pre-filtration but disappeared post-filtration, the locus is a candidate genetic locus of a control target trait. Full-length sequence of the genes is acquired by comparison with the genomes of the parents, target genes are cloned by using PCR amplification, and functions of the target genes cloned are verified on the basis of genetic complementation assay. The method clones directly the genes themselves, thus avoiding isolation and identification of microbial subcultures. |
