Construction and identification of luciferase reporter gene vectors containing two haplotypes DNA of human NLRP1 gene promoter region

This study is aimed to construct a luciferase reporter gene vector containing two different haplotypes DNA on SNP rs878329 of human NLRP1 gene promoter region.The 476 bp DNA fragments of NLRP1 promoter region including the SNP rs878329 were obtained by polymerase chain reaction(PCR) based on human genome DNA from the subjects with the GG/CC genotypes.Then the DNA fragments were digested by restriction endonucleases KpnⅠ and BglⅡ.After fragment recovery,those fragments were ligated to pGL3-promoter vectors,which had digested by restriction endonucleases KpnⅠ and BglⅡ as well,to construct recombinant plasmids pGL3-promoter-G and pGL3-promoter-C.By sequencing,we confirmed that the two recombinant plasmids(pGL3-promoter-G and pGL3-promoter-C) containing two haplotypes DNA of human NLRP1 promoter region were obtained.These expression vectors constructed in our study will be important tools for exploring the relation between the SNP rs878329 and the expression of NLRP1 gene.