乳酸乳球菌NICE系统食品级分泌表达载体pRNV48的构建及铜绿假单胞菌融合外膜蛋白的表达

Construction of a Food-Grade NICE System Vector pRNV48 in Lactococcus lactis and the Expression of Fusion OprF/H from Pseudomonas aeruginosa

乳酸菌NICE系统是目前较理想的可控制蛋白质生产的食品级诱导表达系统,而乳酸菌分泌表达异源蛋白比细胞质表达更优越。本研究将以pVE5524为模板PCR扩增的1.5 kbSPUsp45-NucA-CWAM6-t1t2基因克隆到食品级细胞内诱导表达载体pRNA48上,构建成食品级细胞壁锚定表达载体pRNV48,与宿主菌L.lactisNZ9000共同构成乳酸乳球菌食品级细胞壁锚定诱导表达系统。为检测该系统的功能,将铜绿假单胞菌融合外膜蛋白基因OprF/H克隆进pRNV48中,并检测了OprF/H的表达量和免疫原性。

The food - grade nisin - controlled expression （NICE） system of lactic acid bacteria （LAB） is the overproduction and inducible expression system. The heterologous proteins secreted out of Lactococcus lactis is better than cytoplasmic production. The 1.5kb SP Usp45 - NucA - CWA M6 - tlt2 gene from the plasmid pVE5524 was cloned into a food - grade cytoplasmic inducible expression vector pRNA48 and named as pRNV48 which was used to construct the L. lactis food -grade inducible cell wall anchored expression system together with the L. Lactis NZ9000. To validate the expression of heterologous in the recombinant vector, OprF/H, a fusion gene from outer membrane protein F and H of Pseudomonas aeruginosa, was cloned into pRNV48, and the cell wall anchored OprF/H and its immunogenesity were inspected.