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称为雄激素(AGH)的多肽激素控制的。陆生等足动物Armadillidium uggare AGH(ARV-AGH)的一级结构最终确定为异源二聚体糖蛋白。在成熟的ARV-AGH中，一个N-连接的糖链部分被发现是生物活性所必需的。因此，在本研究中，利用杆状病毒表达系统获得了含有糖链部分的重组AGH。用含有ARV-AGH基因插入片段的重组杆状病毒表达载体感染昆虫细胞Sf9，获得了重组的ARV-AGH。在用抗ARV-AGH抗体进行Western印迹分析中，检测到糖基化的重组ARV-AGH的免疫活性条带。最后，从罗氏沼虾中克隆了编码AGH样肽的基因(MAR-IAG)，并用与重组ARV-AGH相同的方法表达了含有糖基的重组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.