人内皮抑素与IL3信号肽融合蛋白真核表达载体的构建

Construction of the eukaryotic expression vector expressing the fusion protein of human endostatin protein and IL3 signal peptide

AIM To construct the eukaryotic expression vector which expresses the fusion protein of IL_3 signal peptide and human endostatin (endo)protein. METHODS IL_3 signal sequence and human endostatin gene were amplified with PCR and spliced with overlap extension PCR. The IL_3-endo cDNA was cloned into mammalian expression vector pcDNA3.1 ( ). RESULTS The chimeric gene, about 701bp, which included IL_3 signal sequence and entire human endostatin gene, was successfully spliced and cloned into eukaryotic expression vector pcDNA3.1 ( ) downstream from the CMV promoter. The data of nucleotide sequence determination showed that the chimeric cDNA sequence was the same as reported sequence, and in one ORF. CONCLUSION The recombinant vector containing chimeric gene of IL_3-endostatin can express IL_3-endostatin protein in mammalian cell and secrete to extracellular matrix from cells. The success of constructing this vector laid the foundation for studying tumor gene therapy by endostatin.