猪吲哚2,3-二氧化酶介导人血清对猪血管内皮细胞杀伤作用的研究-study on the killing effect of human serum on porcine vascular endothelial cells mediated by porcine indole 2,3 - dioxygenase.docxVIP

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猪吲哚2,3-二氧化酶介导人血清对猪血管内皮细胞杀伤作用的研究-study on the killing effect of human serum on porcine vascular endothelial cells mediated by porcine indole 2,3 - dioxygenase.docx

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猪吲哚2,3-二氧化酶介导人血清对猪血管内皮细胞杀伤作用的研究-study on the killing effect of human serum on porcine vascular endothelial cells mediated by porcine indole 2,3 - dioxygenase

华中科技大学博士学位论文华中科技大学博士学位论文 PAGE 7 PAGE 7 Effect of pocine IDO in Triggering Xenogeneic Cytolytic Activity of Human Serum Against Porcine Endothelial Cells Institute of Pediatric Surgery Tongji Hospital, Tongji Medical College Huazhong University of Science Technology PhD Candidate: Chao Chen Supervisor: Prof. Ming-Fa Wei Abstract Part Ⅰ. Bioinformatics prediction, molecular cloning and characterization analysis of porcine IDO Objective To confirm the existence of IDO gene in porcine, and to clone the novel gene followed by the molecule structure properties and expression pattern analysis. Methods The porcine mRNA sequence homologous to human IDO were obtained from GenBank database by bioinformatics method. With RT-PCR，the gene was cloned from porcine endothelial cell line and confirm the accuracy of the nucleic acid sequence, then followed the expression pattern of the gene. Chromosomal localization was performed by the way of somatic cell hybrid panel(SCHP). The three-dimension structure model of porcine IDO was built referring to the tertiary structure of human IDO using biological sequence analysis softwares and database. Results Porcine IDO was identified by sequencing. The nucleotide sequences were confirmed as a novel gene after submitted to Genbank. Porcine IDO was expressed in lung、 thymus、epididymis and anterior chamber with basic level, however in PBMC with high expression level. Chromosomal localization is yet to be done.The three-dimension structure model of porcine IDO is similiar to human IDO. Conclusion Integrated application of molecular biology techniques and bioinformatics method will conduce the prediction and probation of novel genes. Identification of the structure information of porcine IDO is essential to further investigate the immunologic function of the gene. Key words expressed sequence tag；IDO；bioinformatics； porcine endothelial cell； somatic cell hybrid panel Part Ⅱ. Construction, expression and purification of porcine pcD