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Functional expression of mouse insulin-like growth factor-I with food-grade vector in Lactococcus lactis NZ9000
Authors:Gao G  Qiao J-J  Yang C-H  Jiang D-Z  Li R-Q  Su J-J  Xu H-J  Zhang X-M  Bai Y-L  Qiao M-Q
Institution:The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, College of Life Sciences, Nankai University, Tianjin, China.
Abstract:Aims: To functionally express the recombinant mouse insulin‐like growth factor‐I (rtmIGF‐I) in Lactococcus lactis NZ9000 with a food‐grade vector. Methods and Results: The rtmIGF‐I encoding sequence was inserted into secreted food‐grade vector pLEB688 and transformed into L. lactis NZ9000. The expression of the recombinant protein rtmIGF‐I was confirmed by tricine‐SDS‐PAGE analysis and Western blot. The concentration of this recombinant protein was 3 mg l?1 in the medium fraction. Further experiment demonstrated that the recombinant protein was biologically active and promoted NIH3T3 cell proliferation in a concentration‐dependent manner. Conclusions: The rtmIGF‐I was expressed in L. lactis and located into the medium fraction. The optimal final concentration which could promote NIH3T3 cell proliferation after incubation was 100 ng ml?1. Significance and Impact of the Study: The rtmIGF‐I was functionally expressed in L. lactis NZ9000 with a food‐grade vector. Thus, the recombinant L. lactis NZ9000 could act as a host for the production of rtmIGF‐I for further study. The recombinant strain could serve as an IGF‐I delivery system.
Keywords:biologically active  food‐grade  Lactococcus lactis  recombinant mouse IGF‐I
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