深海沉积物来源宏基因组文库的构建与生物活性物质的功能筛选-微生物学专业论文.docxVIP

深海宏基因组文库的构建与生物活性物质的功能筛选Abstract 深海宏基因组文库的构建与生物活性物质的功能筛选 Abstract Deep。sea accounts for 49％of the earth’S surface SO as to be the earth’S largest bio-habitat areas．Biomining the biological(microbial)genetic resources shows the great practical and scientific importance。Typical deep-sea environment is characterized by low temperature(5(2)and high pressure(up to 1 1 0MPa)．It is estimated that there may be less than 0．0 1％cultured microbes under this extreme circumstances．Metagenomic approach can be used to overcome at least partially the bias，gain access to new scientific discoveries and genetic resources． Three metagenomic libraries with deep-sea sediments samples through previously enrichment methods we佗constructed．Functional screening of antitumor(cytotoxic)，antibacterial and antivirus activity from fermentation products originated from three enriched metagenomic libraries and two previously constructed environmental deep-sea metagenomic libraries clones was performed． Chemical diversity analysis showed that fermentation products from deep—sea environmental metagenomic libraries clones were majorly from E．coli background． Functional screening resulted in indentification several antitumor(cytotoxic)mixed clones and three antiviral monoclones． 1 6 color-producing clones which showed antifungus and or antitumor(cytotoxic) were found in three enriched deep-sea metagenomic libraries．Six cyclo dipeptides and three indol derivatives compounds including a new structure and cytotoxic(ICso aroud 25 I_tg／m1)(2-hydroxyphenyl)(di-1H-indol一3-y1)acetic acid were isolated and characterized from two clones 5C 11 and“F6． Thr01．tgh full lens sequencing of ten color-producing clones，one aromatic hydrocarbon degradation gene cluster and two phenol degradation gene clusters were identified．Transposon mutation analysis，HPLC fingerprints analysis and cytotoxic detection results demonstrated that these clusters a佗closely related with cytotoxic activity of second metabolism products from thes