Construction and space conformation analysis of the fusion gene of human IgG3 upper hinge region and p53 tetramerization domain / 细胞与分子免疫学杂志

ABSTRACT

Aim To construct the fusion gene of human IgG3 upper hinge region and p53 tetramerization domain and to analyze its space conformation. Methods The fusion gene was obtained by recursive polymerase chain reaction (R-PCR),and was cloned into vector pUC19. The positive clone was selected and sequenced with PE310 auto-sequencer. The space conformation of the fusion gene expression product was predicted by using computer program Antheprot. Results Restriction endonuclease digestion confirmed that the fusion gene has been inserted correctly into the vector. The result of sequencing showed that the fusion gene is identical with designation. Analyzing with antheprot program showed that the fusion gene expression product could auto-assembly into a tetramer with four long and flexible linkers. Conclusion Successful construction of the fusion gene mentioned above laid the foundation for further preparation of multivalent gene engineering antibody.