文档介绍：
sFGFR2IIIb 的原核表达及其对 H1648 细胞
增殖的抑制作用
朱保伟,孙莉,王宜,曾威红,李珍,汪炬***
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(暨南大学生命科学技术学院,生物医药研究院/细胞生物学系,广州 510632)
摘要:目的:克隆 sFGFR2IIIb 胞外段,原核表达并纯化。方法:将目的 sFGFR2IIIb 胞外段
基因转入 pET3c 载体,转化进入 BL21 工程表达菌株中,包涵体形式表达胞外段。经凝胶层
析法复性、肝素亲和层析纯化,得到高纯度的目的蛋白。然后 SDS-PAGE 电泳鉴定纯度,
CCK-8 法验证生物活性。结果:成功克隆获得 660bp 大小的人 sFGFR2IIIb 基因片段,诱导
表达包涵体,K-8 法证实该胞外段能明显抑制人
肺非小细胞癌细胞(H1648)的增殖,结果显示在 320ng/ml 时抑制作用最为明显。结论:成功
获得纯度高的 sFGFR2IIIb 胞外段蛋白,经实验验证具有较好的生物活性,这为进一步的研
究奠定了基础。
关键词:sFGFR2IIIb;原核表达;复性纯化;活性
中图分类号:Q786
Prokaryotic expression of sFGFR2IIIb and its inhibitory
effect on H1648 cell proliferation
ZHU Baowei, SUN Li, WANG Yi, ZHENG Weihong, LI Zhen, WANG ju*
(Biological Medicine Research Institute,College of life science and technology, Jinan University,
GuangZhou 510632)
Abstract: Objective:Cloning,expression ang purification sFGFR2IIIb. Method:The gene of
sFGFR2IIIb extracellular domain was transferred to pET3c vector and then transformed into BL21
engineered to express the strains in the form of inclusion bodies ectodomain. By gel
chromatography refolded heparin affinity chromatography purification to obtain a high purity of
the target protein. Then identified by SDS-PAGE purity, CCK-8 method to verify biological
activity. Results: essfully cloned 660bp size sFGFR2IIIb gene fragment, induced inclusion
bodies, refolding of the target protein obtained after purification