转胸腺素基因螺旋藻的表达及免疫增强活性研究
Expression of Thymosin α1 (Tα1) in spirulina platensis and its function of immunity enhancement as feed additive
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摘要: 胸腺素α1(Tα1)是具有免疫增强作用的28肽,通过基因工程技术构建获得了转Tα1)基因螺旋藻,分别得到了1~4个串联目的基因的表达株,ELISA和HPLC检测结果表明:最高表达量可达藻细胞可溶性蛋白的7%,通过饲喂鳗鱼苗试验表明能有效提高鳗鱼苗体内的胸腺素含量,增强鳗鱼苗的免疫力。免疫增强蓝藻作为鱼类饲料添加剂适口性好,每公斤饲料适宜添加量为0.5~1.0 g,可望成为水产养殖的新型免疫增强饵料藻。Abstract: Thymosin α1 (Tα1) is a 28 amino acid peptide, which as immune system enhancer.It had been expressed and purified in a kind of blue green algae, Spirulina platensis.The expression would produce a valuable and cheaper bio—product, because it omitted an expensive purifying process and can be used as an oral medicine or feed additive directly.In this study, A cluster expression system was designed and a gene integration platform system for Spirulina platensis was constructed.ELISA and HPLC analysis was used to compare the difference of expression amount among the 1 to 4 Tα1) gene cluster in transgenic Spirulina platensis.The results showed that the cluster expression was effective and the maximum amount of Tα1 reached 7% of total protein.The immunity enhancement by the transgenic Spirulina platensis was demonstrated by feeding Anguilla japnica as feed additive.Tα1 contents in the tested tissues and organs of the black fry increased after fed with transgenic algae.The transgenic immunity-stimulated Spirulina platensis was the feasible additive of formulated feed for farming fish and its additive dosages was about 0.5—1.0 g·kg-1.