Functional expression of mouse insulin-like growth factor-I with food-grade vector in Lactococcus lactis NZ9000
Functional expression of mouse insulin-like growth factor-I with food-grade vector in Lactococcus lactis NZ9000
复制标题
小鼠胰岛素样生长因子-I与食品级载体在乳酸乳球菌NZ9000中的功能表达
DOI：
10.1111/j.1472-765x.2012.03222.x
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发表时间：
2012-05-01
影响因子：
2.4
通讯作者：
Qiao, M. -Q.
中科院分区：
文献类型：
--
作者：
Gao, G.;Qiao, J. -J.;Qiao, M. -Q.
目的：利用食品级载体在乳酸乳球菌NZ9000中表达重组小鼠胰岛素样生长因子-I(rtmIGF-I)。方法与结果：将rtmIGF-I编码序列插入食品级分泌型载体pLEB688，转化乳酸乳球菌NZ9000。经Tricine-SDS-PAGE和Western印迹分析证实重组蛋白rtmIGF-I在大肠杆菌中得到了表达。重组蛋白的浓度为3 mg L(-1)。进一步的实验证明重组蛋白具有生物学活性，并以浓度依赖的方式促进NIH3T3细胞的增殖。结论：rtmIGF-I在乳酸乳杆菌中得到了表达，并定位于中间组分。促进NIH3T3细胞增殖的最佳终浓度为100 ng ml(-1)。本研究的意义和影响：利用食品级载体在乳酸乳杆菌NZ9000中实现了rtmIGF-I的功能性表达。因此，重组乳酸乳杆菌NZ9000可以作为rtmIGF-I生产的宿主进行进一步的研究。重组菌株可作为IGF-I的载体系统。
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.