关键词: 核心肽, 精氨酸聚合体, 重组融合蛋白质类, 载体蛋白类, 聚合酶链反应, 方法

Abstract: Objective To construct the prokaryotic expression vector of the fusion protein composed of protein transduction domain-eight arginine (8R) and MUC1 core peptides, and to express and purify the 8R-MUC1 core peptide fusion protein which should possess biological activity. Methods Two tandem repeat sequence of MUC1 core peptides were gotten from HSP65-MUC1-pET28a plasmid by PCR and cloned into pMD18-T vector. The T vector was digested by endonuclease for releasing the fragment of two repeats of MUC1 core peptides which was used for constructing six tandem repeat sequence of MUC1 core peptides by ligation. The fragment of six repeats of MUC1 core peptides was cloned into pET26b⁺ plasmid, which contained 8R, for constructing expression vector of the fusion protein composed of 8R and MUCPT (8R-MUCPT). The E.coli BL21(DE3) transformed with 8R-MUCPT-pET26b⁺ were induced by IPTG for expression of 8R-MUCPT fusion protein. The protein was purified by Ni²⁺ affinity chromatography. Results The prokaryotic expression vector, 8R-MUCPT-pET26b⁺, was constructed with 8R and six tandem repeat sequence of MUC1 core peptides. The fusion protein of 8R-MUCPT was expressed in BL21(DE3). The purity of the fu sion protein was 95.5% after purified by Ni²⁺ affinity chromatography. Conclusion The prokaryotic expression vector of 8R-MUC1 core peptide fusion protein has been constructed, and the fusion protein with biological activity has been successfully expressed and purified.

Key words: core peptides, poly-arginine, recombinant fusion proteins, carrier proteins, polymerase chain reaction, methods