推荐回答(3个)
E.coli的乳糖操纵子是原核生物基因表达调控的典型例子.
1�乳糖操纵子的结构
大肠杆菌的乳糖操纵子含Z、Y及A三个结构基因,分别编码β-半乳糖苷酶、透酶、乙酰基转移酶,此外还有一个操纵序列O、一个启动序列P及一个调节基因Ⅰ。Ⅰ基因编码一种阻遏蛋白,后者与O序列结合,使操纵子受阻遏而处于转录失活状态。在启动序列P上游还有一个分解(代谢)物基因激活蛋白CAP结合位点,由P序列、O序列和CAP结合位点共同构成LAC操纵子的调控区,三个酶的编码基因即由同一调控区调节,实现基因产物的协调表达。
2�阻遏蛋白的负性调节
在没有乳糖存在时,乳糖操纵子处于阻遏状态。此时,Ⅰ基因列在P启动序列操纵下表达的乳糖阻遏蛋白与O序列结合,故阻断转录启动。阻遏蛋白的阻遏作用并非绝对,偶有阻遏蛋白与O序列解聚。因此,每个细胞中可能会有寥寥数分子β半乳糖苷酶、透酶生成。
当有乳糖存在时,乳糖操纵子即可被诱导。真正的诱导剂并非乳糖本身。乳糖经透酶催化、转运进入细胞,再经原先存在于细胞中的少数β -半乳糖苷酶催化,转变为别乳糖。后者作为一种诱导剂分子结合阻遏蛋白,使蛋白构型变化,导致阻遏蛋白与O序列解离、发生转录,使β-半乳糖苷酶分子增加 1000倍。
3�CAP的正性调节
分解代谢物基因激活蛋白 CAP是同二聚体,在其分子内有DNA结合区及cAMP结合位点。当没有葡萄糖及cAMP浓度较高时,cAMP与CAP结合,这时CAP结合在乳糖启动序列附近的CAP位点,可刺激RNA转录活性,使之提高50倍;当葡萄糖存在时,cAMP浓度降低,cAMP与CAP结合受阻,因此乳糖操纵子表达下降。
由此可见,对乳糖操纵子来说 CAP是正性调节因素,乳糖阻遏蛋白是负性调节因素。两种调节机制根据存在的碳源性质及水平协调调节乳糖操纵子的表达。
4�对调节机制的解释
大肠杆菌根据碳源性质选择代谢方式。
倘若有葡萄糖存在时,细菌优先选择葡萄糖供应能量。葡萄糖通过降低 cAMP浓度,阻碍cAMP与CAP结合而抑制乳糖操纵子转录,使细菌只能利用葡萄糖。
在没有葡萄糖而只有乳糖的条件下,阻遏蛋白与 O序列解聚,CAP结合cAMP后与乳糖操纵子的CAP位点,激活转录,使得细菌利用乳糖作为能量来源。
乳糖操纵子包括调节基因、启动基因、操纵基因和结构基因。乳糖操纵子模型是两重调控:乳糖负调控,CAP正调控。
当培养基没有乳糖时,阻遏蛋白与操纵基因结合,从阻止了RNA聚合酶与DNA的结合,转录停止。
但是有乳糖时,在β—半乳糖苷酶的作用下,乳糖转变为异构乳糖从而与阻遏蛋白结合,使之构象发生改变,不能与操纵基因序列结合,从而DNA可转录。这就是乳糖操纵子的负调控。
当培养基有葡萄糖且充足时,CAMP水平很低,并且CAMP很少与CAP结合,导致RNA聚合酶无法高效结合到DNA,DNA转录低水平。
但是葡萄糖含量少时,CAMP水平很高,CAP很容易结合CAMP,形成复合物结合DNA,增强RNA聚合酶结合效率,DNA转录并翻译。这就是乳糖操纵子中CAP的正调控。
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