四、細胞核轉錄因子的分離和鑑定
信號轉導的結果是細胞核內一些轉錄因子同DNA上某些特定的序列結合,調節轉錄活性和基因的表達水平。因而分離鑑定細胞內核轉錄因子在信號轉導研究中具有獨特的意義。根據核轉錄因子可與一些特定的核甘酸序列結合的特點,將其特定的核苷序列標記上同位素,來鑑定信號轉導中的核轉錄因子。核轉錄因子具有一些特徵性的結構域,即亮氨酸拉鏈(lucine zipper)、鋅指結構(zinc finger)和螺旋-轉角-螺旋(helix-turn-helix)結構。並且受用足跡法(foot printing)分析新發現核轉錄因子結合DNA的位置和核甘酸序列。
隨著分子生物學核技術的發展,信號轉導研究取得很大進展。採用基因克隆技術發現了越來越多的信號蛋白分子;嵌合分子的構建及基因轉染等技術的應用為細胞因子的信號轉導研究提供了有效的手段;點突變、肽競爭等技術使信號蛋白中某些功能區的作用的研究變得更為精確。
(夏海濱 金伯泉)
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