原核生物基因表达调控预案.ppt

乳糖操纵子所受的负调控 (negative regulation) 负调控系统中主要的作用因子是阻遏蛋白(Repressor )，诱导物乳糖(lactose)的结合可使之失活。 Repressor and RNA polymerase bind at sites that overlap around the transcription startpoint of the lac operon. Operator An inducer (such as lactose) is a small molecule that triggers gene transcription by binding to a regulator protein. A corepressor is a small molecule that triggers repression of transcription by binding to a regulator protein. In the absence of an inducer, the genes are not transcribed, because repressor protein is in an active form that is bound to the operator. When an inducer is added, the repressor is converted into an inactive form that leaves the operator. · An inducer functions by converting the repressor protein into an inactive form. · Repressor has two binding sites, one for the operator and another for the inducer. · Repressor is inactivated by an allosteric变构的 interaction in which binding of inducer at its site changes the properties of the DNA-binding site. 诱导物inducer 乳糖是lac操纵子转录的活性诱导物，人们发现一个合成的、结构上类似于乳糖、不能被β-半乳糖苷酶水解的β-半乳糖苷异丙基硫代半乳糖苷（isopropyl thiogalactoside：IPTG）起着lac操纵子的一个诱导物的作用，所以IPTG常用于诱导含有使用了lac启动子的质粒载体的细菌中的重组蛋白的表达。 lac 操纵子受阻遏蛋白调控。在没有诱导物（乳糖或IPTG）存在时，阻遏蛋白与lac 操纵子DNA序列结合得非常紧密，lac 基因不能进行转录。当诱导物存在时，诱导物与阻遏蛋白相互作用，使得阻遏蛋白变构，对lac 操纵基因的亲和性为单独阻遏蛋白的亲和性的千分之一。 lac 操纵子一般处于负转录调控状态，即在诱导物缺乏的条件下，抑制RNA聚合酶的转录。培养基中有乳糖时，诱导lac 操纵子的正常表达  乳糖操纵子负调控总结 控制区基因突变造成的影响： lacI 或 lacO， lacP的突变影响乳糖操纵子中三个结构基因的表达： lacI- ，阻遏蛋白失活， lacZ、Y、A永久表达，即组成型表达(constitutive expression) lacOc，此突变造成lacO序列不再被阻遏蛋白识别结合，故下游结构基因组成型表达 lacIs ，阻遏蛋白变成超阻遏物, 不能与诱导物结合而永久结合在lacO上, 乳糖操纵子永久关闭 it metabolizes the glucose and represses the use of lactose. 这一现象涉及到乳糖操纵子的正调控系统。 只有葡萄糖时，利用葡萄糖；只有乳糖时，利用乳糖。 So when E. coli finds both glucose and lactose in the medium ? 乳糖操纵子所受的正调控Positive regulation 正调控系统中主要的作用因子是cAMP-CAP ，葡萄糖通过降低细胞内的cAMP的含量来抑制它的作用。 Catabolite activator protein (CAP，分解代谢产物激活蛋白) is active only in the presence of cyclic AMP. 乳糖操纵子所受的正调控Positive regulation Activator(CAP) 活性由其信号变构调控