[Objective]In order to improve expression vector of L.lactis at food grade and widen application range of original system.[Method] Taking pNZ8149 as basis,the promoter PnisA of pNZ8149 was inserted L.lactis MG1363 and...[Objective]In order to improve expression vector of L.lactis at food grade and widen application range of original system.[Method] Taking pNZ8149 as basis,the promoter PnisA of pNZ8149 was inserted L.lactis MG1363 and SPusp45 of unknown secretory protein in downstream.Through PCR technology,specific primers were used to delete restriction sites between promoter sequence and signal peptide gene sequence and ensure better distance between SD sequence and start codon to construct secreting expression vector pNZS.The reporter gene gus was recombined into multiple cloning site of pNZS to construct pNZS-gus and L.lactis was transformed by electroporation.10 ng/ml nisin was used for induction culture,then culture solution was conducted GUS staining test.[Result]The new constructed L.lactis N3900/pNZS-gus system could express active GUS protein and GUS protein could be secreted out of cell.[Conclusion]The successful construction of this system lays foundation for secretion expression study of protein and oral vaccine research.展开更多
基金Supported by Doctoral Fund of Jilin Agricultural University(20070193005)~~
文摘[Objective]In order to improve expression vector of L.lactis at food grade and widen application range of original system.[Method] Taking pNZ8149 as basis,the promoter PnisA of pNZ8149 was inserted L.lactis MG1363 and SPusp45 of unknown secretory protein in downstream.Through PCR technology,specific primers were used to delete restriction sites between promoter sequence and signal peptide gene sequence and ensure better distance between SD sequence and start codon to construct secreting expression vector pNZS.The reporter gene gus was recombined into multiple cloning site of pNZS to construct pNZS-gus and L.lactis was transformed by electroporation.10 ng/ml nisin was used for induction culture,then culture solution was conducted GUS staining test.[Result]The new constructed L.lactis N3900/pNZS-gus system could express active GUS protein and GUS protein could be secreted out of cell.[Conclusion]The successful construction of this system lays foundation for secretion expression study of protein and oral vaccine research.
