深海宏基因组文库克隆子发酵产物的生物活性筛选

Bioactivity Screening of Fermentation Crude Extracts from Deep Sea Metagenomic Library Clones

目的:对前期构建的深海宏基因组文库克隆子发酵产物进行生物活性筛选。方法:利用CCK8法及琼脂扩散法和双层琼脂法进行了细胞毒活性及抗菌活性筛选,利用荧光定量PCR法进行了抗乙肝病毒活性筛选。结果:筛选发现3个具有细胞毒活性的混合克隆子。HPLC指纹图谱分析表明这3个混合克隆子具有与大肠杆菌宿主对照不同的指纹图谱,其活性与出现的指纹峰成分可能相关。结论:研究结果提示对深海宏基因组文库克隆子发酵产物进行化学筛选为常规的生物活性功能筛选之外的有意义途径。

Objective： Detect bioactivities of fermentation crude extracts originated from previously constructed deep sea metagenomic libraries. Methods： Functional screening of antitumor（cytotoxic） ,antibacterial and anti HBV activity from fermentation crude extracts was performed by CCK-8 method, agar diffusion method, double agar method and real time PCR method. Results： No antibacterial and anti HBV clones were identified and three mixed clones showed good cytotoxic activity. HPLC analysis of these three mixed clones fingerprints indicated they possess obviously different fingerprints with E. coli host which could be useful to next isolation of cytotoxic compounds from deep sea metagenomic library. Conclusion： These results indicated that chemical screening could be potential way for screening bioactivity substance from deep sea metagenomic libraries besides functional screening and homology screening.