简介在一段时间内同时检测几种不同信号的输出尤其在研究蛋白或化合物对细胞生长或基因表达作用方面很有帮助。在本应用指南里,我们利用SoftMax? Pro 7版本软件同时来检测细胞生长(光吸收)和蛋白表达(荧光)。在细菌中,lac操纵子是一种由编码?-半乳糖苷酶的启动子调控的基因群。将一个蛋白表达序列插入到含有lac操纵子的质粒中并转染细菌,可使这一转化细菌来表达感兴趣的蛋白。通常情况下,lac启动子是被变构抑制的,但是在异丙基-?-D-1-硫代半乳糖苷(IPTG)存在下,阻遏物从启动子序列释放引起感兴趣蛋白的的表达。相反,如果IPTG过量,细胞会转变为优先将胞内物质用于蛋白表达并且细胞生长也会受到阻碍。通过同时检测细胞密度和蛋白表达,我们展示了大肠杆菌(E.coli)是如何响应系列稀释的IPTG的。
Gene expression studies are important for understanding generegulation, deducing cell signal cascades, etc. More broadly, thesestudies are often used to understand disease development, as specificgenes are commonly up regulated in clinical pathologies. Thesestudies can be paired with the application of a mechanical stimulus as well for example, endothelial cells can be exposed to a defined fluid flow (physiological or pathological) in order to understand the possible mechanisms for atherosclerosis development.