mlrC基因异源表达系统的构建

Microcystins (MCs) are the most toxic and frequently encountered cyanobacterial toxins. Biodegradation is an effective method for removing MCs. Novosphingobium sp. Construction of a heterologous expression system for mlrC geneTHN1, isolated from Lake Taihu., can degrade MCs effectively, while its mlrC gene is deemed to regulate MCs-biodegradation process. Construction of a heterologous expression system (HES) for mlrC gene provides experimental basis and technical support for elucidating the function of mlrC and mlrC-encoded enzyme (MlrC) characteristics. Through PCR amplification, enzymatic digestion, combination with vector, transformation into heterologous host and SDS-PAGE, this study succeed in constructing a HES for THN1-mlrC and expressing MlrC by induction. This success supports downstream research. Moreover, this study optimized the reaction conditions for vector- combination experiments, causing a simplified procedure for HES construction and optimized the temperature condition for induced expression of mlrC, and also gave suggestions to some concerns during experiments. These may be helpful to other related studies.