The invention discloses a nocardiopsis dassonvillei lasso peptide biosynthetic gene cluster and a cloning and expression method thereof and belongs to the technical field of biology. The lasso peptidebiosynthetic gene cluster of a Nocardiopsis sp.Y469 strain is obtained through genome excavation, and a plurality of recombinant systems of key genes of the lasso peptide biosynthetic gene cluster are constructed by using RecE/T cloning, Red alpha-beta recombination and indirect conjugative transfer molecule genetic operation, so that heterologous expression is realized in an engineering strain,and differential products are obtained. The antibacterial activity of the product is preliminarily screened, and it is found that the product has bacillus subtilis inhibition activity. By adopting thelasso peptide biosynthetic gene cluster and the cloning expression scheme thereof, a new idea and a new genetic material are provided for excavation of natural products of polar microorganisms and development and application of lasso peptides.