引物应用策略:基于2对古菌16S rRNA基因通用引物对环境样品古菌多样性分析

摘要

[Obiective]Finding a series of archaeal 16S rRNA gene universal primer applied strategies to detect complex microbial diversity in environmental samples, especially with rapidly development of next generation sequencing technology challenge.[Methods]We used Oligocheck soft to simulate two pairs of archaeal 16S rRNA gene universal primers with RDP (Ribosomal database project) database 16S rRNA gene sequences matching percentage.In succession,the sediment sample was constructed for two clone libraries by using two pairs of archaeal 16S rRNA gene universal primers.[Results]The soft simulation matched percentage result suggests that primer f109/r958 is better than primer f21/r958.This result is consistent with RFLP and diversity index analyses by two clone libraries.[Conclusion]Multi-primers and properly primer are used to recovery environmental microbiology diversity, which will be advanced in environmental microbial resolution.%[目的]找到适宜的16S rRNA基因通用引物应用策略,应对复杂环境微生物多样性调查,尤其目前高速发展的高通量测序技术带来的巨大挑战.[方法]用Oligoeheck软件分别将两对应试的古菌16S rRNA基因通用引物与RDP(Ribosomal database project)数据库中古菌16S rRNA基因序列进行匹配比对.用两对应试引物分别构建海洋沉积物样品的古菌16S rRNA基因文库.[结果]软件匹配结果显示引物f109/r958与目的基因的匹配程度高于引物f21/r958.该结果与古菌16S rRNA基因文库RFLP分析、古菌多样性指数分析结果相吻合.数据还表明,2对引物的综合文库能更好满足该沉积物样品的古菌多样性分析.[结论]选用与数据库中目的基因匹配性高的通用引物和多个引物的联合使用,可以有效提高环境样品微生物多样性调查的分辨率.