Immunoglobulin A (IgA), a major serum immunoglobulin and a predominant antibody in the external secretions that bathe mucosal surfaces, plays key roles in immune protection. Some pathogenic bacteria including Haemophilus influenzae and Neisseria meningitides, however, produce a protease called IgA1 protease to impair the function of IgA1. The iga mRNA is initially translated into a large precursor containing four distinct domains: a 31-amino acid signal peptide which leads the precursor to the periplasmic space, an 105-kDa protease domain which cleaves host IgA1 molecule, a £]-domain responsible for autotransportation of the protease domain, and a linker £\-protein between the protease and the £]-domain. The hydrolytic function of the protease and the role of the £]-core had been studied extensively, but the role of the £\-protein has never been studied. Thus this study is designed to reveal the possible functions of £\-protein in the proliferation of lymphocytes. To complete the project, PCR was used to amplify the DNA fragment for £\-protein using iga gene (Gene Bank DQ683357) as template. The fragment spans nucleotide numbers of 1015-1405. The fragment was then cloned into pGEX-2T for expression. Recombinant £\-protein was purified using glutathione -Sepharose column. The purified recombinant protein did not seem to affect the cell growth at the concentration of 1 £gg/ml compared with the medium control or GST control. Interestingly, when the concentration was increased to 5 £gg/ml or 10 £gg/ml, £\-protein seems to enhance the cell growth on the 2nd, 4th and 6th day assays, The results suggested that £\-protein may enhance the cell growth in 4 days.
Identifer | oai:union.ndltd.org:NSYSU/oai:NSYSU:etd-0706108-225547 |
Date | 06 July 2008 |
Creators | Tseng, Chiung-ju |
Contributors | Chen-fu Shaw, Shiping He, hung-tu Huang |
Publisher | NSYSU |
Source Sets | NSYSU Electronic Thesis and Dissertation Archive |
Language | Cholon |
Detected Language | English |
Type | text |
Format | application/pdf |
Source | http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0706108-225547 |
Rights | not_available, Copyright information available at source archive |
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