Preparation of an active recombinant peptide of crustacean androgenic gland hormone, a heterodimeric glycopeptide.

甲壳类动物的性别分化是由一种叫做雄激素腺激素（AGH）的肽激素控制的。陆生等足动物Armadillidium vulgare AGH （Arv-AGH）的初级结构最终确定为异二聚体糖蛋白。在成熟的Arv-AGH中发现一个n -链聚糖片段对生物活性至关重要。因此，本研究利用杆状病毒表达系统制备了含有多糖片段的重组AGH。用含有Arv-AGH cDNA插入片段的重组杆状病毒表达载体感染昆虫Sf9细胞，表达重组Arv-AGH。使用抗Arv-AGH抗体进行Western blot分析，检测到糖基化重组Arv-AGH的免疫反应带。最后，从罗氏沼虾（Macrobrachium rosenbergii）中克隆了一个编码agh样肽（Mar-IAG）的cDNA，并通过与重组Arv-AGH相同的方法表达了含有糖基片段的重组Mar-IAG。

The sex differentiation in crustaceans is known to be controlled by a peptide hormone called androgenic gland hormone (AGH). The primary structure of the terrestrial isopod Armadillidium vulgare AGH (Arv-AGH) was finally determined to be a heterodimeric glycoprotein. An N-linked glycan moiety in the mature Arv-AGH was found to be essential for biological activity. Therefore, in this study, a recombinant AGH with a glycan moiety was produced using a baculovirus expression system. Insect Sf9 cells were infected with a recombinant baculovirus expression vector containing an Arv-AGH cDNA insert and subsequently recombinant Arv-AGH was expressed. In Western blot analysis using an anti-Arv-AGH antibody, immunoreactive band of a glycosylated recombinant Arv-AGH was detected.Finally, a cDNA encoding AGH-like peptide (Mar-IAG) was cloned from the giant freshwater prawn Macrobrachium rosenbergii and subsequently a recombinant Mar-IAG with a glycan moiety was expressed by the same methods as recombinant Arv-AGH.