在 Lactococcus lactis NZ9000 中用食品级载体表达鼠胰岛素样生长因子-I 的功能。

Functional expression of mouse insulin-like growth factor-I with food-grade vector in Lactococcus lactis NZ9000.

机构信息

The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, College of Life Sciences, Nankai University, Tianjin, China.

出版信息

Lett Appl Microbiol. 2012 May;54(5):404-9. doi: 10.1111/j.1472-765X.2012.03222.x. Epub 2012 Mar 29.

DOI:10.1111/j.1472-765X.2012.03222.x

PMID:22332956

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.

摘要

目的

利用食品级载体在乳酸乳球菌 NZ9000 中功能性表达重组鼠胰岛素样生长因子-I（rtmIGF-I）。

方法和结果

将 rtmIGF-I 编码序列插入分泌型食品级载体 pLEB688 中，并转化至乳酸乳球菌 NZ9000。通过三氯乙酸-SDS-PAGE 分析和 Western blot 验证重组蛋白 rtmIGF-I 的表达。该重组蛋白在培养基部分的浓度为 3mg/L。进一步的实验表明，该重组蛋白具有生物活性，并以浓度依赖的方式促进 NIH3T3 细胞增殖。