Functional analyais of defense molecule using transgenic mice

通过细菌重组无菌小鼠结肠上皮细胞中鉴定出的抵抗素样分子β (RELM-β)及其在恶性脑肿瘤1（DMBT-1）中缺失的分子功能。我们利用pCAG载体建立了RELM-β转基因小鼠。经过很长一段时间，开始研究转基因的表达。由于未知的原因，其在结肠中的表达异常微弱。此外，葡萄糖聚糖硫酸钠是小鼠结肠炎的诱导剂，经刺激后，正常小鼠与TG小鼠之间无显著差异。在体外实验中，我们发现了RELM-β的抗菌作用，并仍在试图证实这一功能。重组RELM-β结合到金黄色葡萄球菌的细胞表面并破坏其膜。这种蛋白质以二聚物的形式存在于粪便中。另一方面，我们努力克隆全长DMBT-1并制作其重组蛋白，但最终失败。最近的报告表明，克隆DMBT-1需要将核苷酸从原始序列交换到修饰序列（Caroline End等）。蛋白质表达和纯化41,275（2005）。

The aims of the present study were to analyze molecular function of resistin-like molecule β (RELM-β) and deleted in malignant brain tumors 1(DMBT-1), which had been identified in colonic epithelial cells by bacterial reconstitution of germ-free mice.We established RELM-β transgenic mice using pCAG vector. After a long time and started to investigate expression of the trannsgene. Its expression in the colon was unexpectedly weak due to unknown reasons. In addition, there was no significant difference between control and TG mice after challenge of dextran sulfate sodium, which are well accepted as an inducer of colitis in mice.In vitro experiments, we found antibacterial effects of RELM-β and still try to confirm this function. Recombinant RELM-β bound to the cell surface of Staphylococcus aureus and destroyed its membrane. This protein is present as dimmer in stools.On the other hand, we struggled to clone full-length DMBT-1 and make its recombinant protein and finally failed. The recent report suggests that cloning of DMBT-1 requires exchange of nucleotides from original to modified sequences (Caroline End, et al. Protein expression and purification 41, 275 (2005).