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1.Cloning and Expression of Mtb8.4-Hspx and Hspx-Mtb8.4 from Mycobacterial Tuberculosis 2.FTY720 Immunosuppression for Kidney Transplant Recipients

Author: WuYuMin
Tutor: TanJiYing
School: Lanzhou University
Course: Immunology
Keywords: Mycobacterium tuberculosis Mtb8.4 Hspx Clone Expression Kidney transplant FTY720 Immunosuppressants
CLC: R699
Type: Master's thesis
Year: 2010
Downloads: 40
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Abstract


The purpose build Mycobacterium tuberculosis to label recombinant proteins Mtb8.4-Hspx (MH) and Hspx-Mtb8 (HM) prokaryotic expression vector and expressed in E. coli BL21 (DE3) as a tuberculosis subunit vaccine candidate antigen . Mycobacterium tuberculosis H37Rv gene set of DNA templates by PCR amplification Mtb8.4 (containing Nde Ⅰ and Sac Ⅰ site points), Hspx (containing Nde Ⅰ and Sac Ⅰ site points) gene, the product, respectively, by the Nde Ⅰ and Sac Ⅰ digested with the plasmid pET30a connection construct the recombinant plasmid pET30a-Mtb8 and pET30a-Hspx, and transformed into E. coli DH5α. Extraction recombinant plasmid, Sac Ⅰ and Hind Ⅲ respectively digested the recombinant plasmid with the same restriction enzyme digestion of the PCR product Hspx (containing Sac Ⅰ and Hind Ⅲ bit point) and Mtb8.4 (containing Sac Ⅰ and Hind Ⅲ point) connection, build a recombinant plasmid pET30a-Mtb8 the .4-Hspx and pET30a-Hspx-Mtb8.4, the pET30a-MH and pET30a-HM. Transformed into E. coli BL21 (DE3), to a final concentration of 0.5mmol / L of IPTG to induce protein expression, closing bacteria, ultrasonic disrupter sonicated, centrifuged, PBS and resuspended the cell precipitate, row by SDS-PAGE . The results go MH and HM label fusion protein molecular mass of 28,000 line validation and measured by SDS-PAGE protein electrophoresis. Conclusion to go label Mycobacterium tuberculosis recombinant protein MH and HM successful expression, which laid the foundation for further research on tuberculosis subunit vaccine immunogenicity. Objective To evaluate FTY720 on kidney transplant recipients immune suppression effectiveness and safety. Methods We searched MEDLINE (1966-2010); Cochrane Library, EMBASE (1974-2010); (4th); The National Research Register; Chinese Scientific Journals Database (1989-2010), China Academic Journal Full-text database (1994 - 2010). Results of four studies of 1883 patients met the inclusion criteria. Mycophenolate mofetil (MMF) plus full-dose cyclosporine A (CsA) - (MMF FDC) at 6 months and 12 months, compared to a half dose of FTY720 (5.0 mg) plus cyclosporine FDC] group no significant change [P = 0.32, P = 0.60]. FTY720 (2.5mg) FDC contrast MMF FDC, mortality, graft loss rate and antibody treatment of acute rejection were no significant differences in the confidence interval [RR = 0.29, 95% CI 0.04 to 2.17 no significant difference in the bacterial infection (P = 0.43), cytomegalovirus infection (P = 0.44), urinary tract infection (P = 0.92) (P = 0.26) and retinal edema. Conclusion The present study showed that FTY720 (5.0mg) RDC To BPAR incidence improve CsA half dose, it does not support. FTY720 (2.5mg) FDC in the prevention of rejection, to reduce the of mortality and BPAR incidence of MMF FDC similar. Addition of FTY720 groups compared MMF group can reduce the incidence of the infection but the incidence of bradycardia improve the synergy between the drugs with clinical choose medication should pay attention to, pay attention to individual differences, to do joint reasonable medication.

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CLC: > Medicine, health > Surgery > Urology ( urinary and reproductive system diseases) > Urinary tract and male reproductive system surgery
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