Construction Of Vibrio Alginolyticus Omp Gene Expression Vector And Preparation Of Rabbit Anti Sera Anti Large Yellow Croaker IgM

Large yellow Croaker(Pseudosciaena crocea),one of the China's six most superiority export aquatic product,is the main marincultured fish in china with the highest annual output. The most significant factor that affects large yellow croaker culture is the presence of microbial pathologies. V. alginolyticus has been commonly associated with the epidemic vibriosis which leads to mass mortality of cultured large yellow croaker and causes considerable economic losses. DNA vaccine, a rising prevention and cure means with splendid application foreground, develop very fast in recent years.The aim of the present article is to construct V. alginolyticus Omp gene expression vector and prepare rabbit sera anti-IgM of large yellow croaker which could be useful in correlative studies in future.The serum IgM of healthy large yellow croaker was purified by twice salt out of saturated (NH₄)₂SO₄ and protein A-sepharose affinity chromatography. The productions were examined by SDS-PAGE electrophoresis. The results showed that: the production of protein A-sepharose affinity chromatography had only two bands (heavy chain and light chain) in SDS-PAGE, while the production of twice salt out of saturated (NH₄)₂SO₄ has several other bands; moreover, using protein A-sepharose affinity chromatography to purify IgM is more simple,convenient and fast, so protein A-sepharose affinity chromatography was better than twice salt out of saturated (NH₄)₂SO₄ at the purification of IgM; the molecular weight of heavy chain and light chain of Pseudosciaena crocea IgM were 76 kDa and 28 kDa respectively. Sera anti-IgM of Pseudosciaena crocea had been prepared by repetitious immunize New Zealand rabbits with purified IgM, the titers of...