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| Autoinduction | | |  | The Overnight Express Autoinduction System is designed for high-level protein expression with the pET bacterial expression system without monitoring cell growth.In lDE3 lysogenic hosts, uninduced cells are grown to a high cell density followed by spontaneous induction of T7 RNA polymerase expression without monitoring cell density or adding IPTG. The Overnight Express System 1 contains three components: OnEx™ Solution 1 (induction solution); OnEx Solution 2 (buffering solution); and OnEx Solution 3 (magnesium solution). OnEx Solution 1 is a blend of carbon sources optimized for tightly regulated uninduced growth to high cell density followed by high-level induction by lactose and continued growth. OnEx Solution 2 is a concentrated buffer that maintains pH throughout metabolic acid production and supplies additional nitrogen necessary to support increased protein synthesis. OnEx Solution 3 provides critical magnesium for maximum cell density. Addition of these components to traditional glucose-free media such as LB broth, TB, or animal-free Veggie™ media results in maximum yields of target proteins with the pET system. The Overnight Express Autoinduction System 1 is extremely convenient for routine expression of proteins in multiple cultures and is ideal for high-throughput parallel analysis of protein expression, solubility and purification from multiple expression clones. The kits provide sufficient reagents for 1 liter or 5 liters of media. |
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