冬蝶鱼皮肤型抗冻蛋白的克隆及表达

冬蝶鱼皮肤型抗冻蛋白（ｗ ｆｓ Ａ Ｆ Ｐ）的克隆和表达，是研究抗冻蛋白抗冻机制的重要步骤之一，有助于揭示抗冻蛋白分子结构与活力的关系．用 Ｐ Ｃ Ｒ 法获得 ｗ ｆｓ Ａ Ｆ Ｐ 基因片段，构建了分泌型表达载体 Ｐｌａｃ Ｉ Ｑｐａｒ８ｗ ｆｓ Ａ Ｆ Ｐ．用 Ｗ ｅｓｔｅｒｎ Ｂｌｏｔ 法测其表达水平，氨基酸组成分析及质谱分析证明表达的蛋白与天然蛋白完全一致，并具有天然蛋白相同的活力．

Cloning and Expression of Winter-flounder Skin-type Antifreeze Protein

Winter Flounder Skin type antifreeze protein (wfsAFP) has been cloned and expressed. This is one of the important ways to study the mechanism of antifreeze protein. It helps to reveal the relationship between the structure and activity of antifreeze protein. wfsAFP gene fragment was obtained by PCR, and then constructed into secretory expression vector PlacIQpar8cex. Western blot was employed to test the level of the expressed protein. It was comfirmed, by amino acid analysis and Mass spectroscopy, that the expressed protein was the same as native protein, and the activity was the same as well.