摘要
[目的]对159株蜚蠊肠道内生放线菌的卤化酶等相关基因进行检测和初步分析。[方法]活化蜚蠊肠道内生放线菌,制备各菌株基因组DNA,根据依赖黄素腺嘌呤二核苷酸FADH2的卤化酶基因保守区设计简并引物,通过PCR扩增卤化酶基因片段,TA克隆后进行测序鉴定。在此基础上,对卤化酶阳性菌株进行聚酮合酶PKSⅠ和非核糖体多肽合成酶NRPS等2个次级代谢产物生物合成主要基因进行检测。[结果]159株蜚蠊肠道放线菌有84株含有卤化酶基因,占比52.83%;卤化酶基因阳性放线菌中71株含有PKSⅠ基因,70株含有NRPS,占比分别为84.52%、83.33%。[结论]蜚蠊肠道含有较丰富的卤化酶基因阳性放线菌,可作为未来分离卤化活性物质的微生物资源。
[Objective] To detect and preliminary analysis Halogenated enzymes and other related genes of 159 strains of Periplaneta americana intestinal endophytic actinomycetes at the genetic level. [Methods] After Periplaneta americana intestinal endophytic actinomycetes activated and genomicDNA of each strain prepared,Flavin adenine dinucleotide FADH2-dependent halogenases gene was detected by PCR amplification. Degenerate primers were designed according to their conservative region. Then the gene was cloned into the T vector for sequencing. On the basis of this,polyketide synthase PSKⅠgene and non ribosomal peptide synthetase NRPS gene responsible for producing secondary metabolite were tested in the positive isolates of halogenated enzymes. [Results]84 Periplaneta americana intestinal endophytic actinomycetes were found containing FADH2-dependent Halogenases gene and the positive rate was 52. 83%. Among the halogenase gene-positive actinomycetes,71 actinomycetes contained PKSⅠ gene,70 actinomycetes contained NRPS gene,the positive rate was 84. 52% and 83. 33% respectively. [Conclusion] There are many halogenase gene-positive actinomycetes in Periplaneta americana intestinal,which can be used as a microbial resource for the future to separate halogenated active substances.
出处
《生物技术》
北大核心
2017年第5期466-471,共6页
Biotechnology
基金
广东省公益研究与能力建设项目("基于中药蜚蠊肠道内生放线菌资源筛选药物先导化合物"
No.2016A030303059
"液质联用代谢组学技术筛选中药蜚蠊放线菌抗真菌化合物及其作用机制研究"
No.2017A020211008)
关键词
蜚蠊
放线菌
卤化酶
基因检测
分析
Periplarteta americana, actinomycetes, halogenase, gene detection, analysis
