| Summary: | 碩士 === 國立臺灣海洋大學 === 水產養殖學系 === 95 === IgM is the main immunoglobulin of teleost. Constant region of IgM heavy chain gene of grouper includes Intron 1, Intron 2 and Intron 3 composed of 1395, 117 and 175 base pairs separately, and CH1, CH2, CH3 and CH4 composed of 340, 342, 321 and 392 base pairs separately. The expression of IgM heavy chain mRNA is in spleen, head kidney and kidney. It is also expressed in mucus tissues including skin, gill and intestine and can strengthen the humoral immune response for teleost. But liver, muscle and brain do not express IgM cDNA. IgM mRNA was expressed the seventh day after fertilization the fifth day after hatched. Grouper IgM-CH cDNA was isolated and cloned into expression vector pET-15b. Recombinant protein IgM-CH was induced with IPTG and purified by His-Tag affinity chromatography. The recombinant IgM-CH was used to immunize rabbit. The rabbit anti-IgM-CH antiserum can be used to detect the heavy chain of immunoglobulin of various kinds of groupers, and a 45 kDa protein which may be a kind of IgM heavy chain homology (isotype) protein. Anti-IgM-CH antiserum can also be used to detect the IgM heavy chain of Rachycentron canadum, Rhabdosargus sarba, Lutjanus erythropterus and Trachinotus blochii.
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