LKB1(Liver Kinase B1) gene is a tumor suppressor gene, which regulates the pathophysiological function of many cells, but the mechanism of LKB1 gene is still unclear. The double transgenic mice with positive homozygous and negative homozygous were screened by PCR amplification technique and gel electrophoresis technique. The expression level of LKB1 gene in smooth muscle cells was detected by Westernblot technique. We has successfully obtained the gene type for the LKB1−/−, CRE+ smooth muscle of LKB1 condition gene knockout mouse after identification, which for the vast number of scientific experimenters provides mouse model to study the function of LKB1 gene in smooth muscle cell and to search for new related disease treatment method batch.