CAJ | 학술논문

本研究从担子菌毛头鬼伞(Coprinus comatus)菌丝中分离获得一条新的28S rRNA序列,序列长度为906 bp(GenBank accession No.GU568178).该序列是我们前期在从毛头鬼伞中克隆一种烟草花叶病毒(TMV)的抗性蛋白基因y3时意外获得的一条非目的条带.将此获得的序列通过NCBI的BLAST,以及与其同源序列进行Clustal w和MEGA聚类分析,证实该序列是28S rRNA,同时还发现毛头鬼伞的系统进化关系比较离散.此外,在这一新28S rRNA与TMV的抗性蛋白基因y3之间发现有两个同源区段有可能是PCR扩增y3基因时出现非目的条带的原因.在这两个同源区段中,其一区段与克隆y3基因时所用的PCR引物之一有较高的相似性,另一区段也是一般PCR引物的类似物.本研究中新28S rRNA序列的获得是PCR扩增中出现非目的条带的新例,该序列的发现及聚类分析的结果有助于真菌基因组学研究及真菌生物分子分类系统的建立.
본연구종담자균모두귀산(Coprinus comatus)균사중분리획득일조신적28S rRNA서렬,서렬장도위906 bp(GenBank accession No.GU568178).해서렬시아문전기재종모두귀산중극륭일충연초화협병독(TMV)적항성단백기인y3시의외획득적일조비목적조대.장차획득적서렬통과NCBI적BLAST,이급여기동원서렬진행Clustal w화MEGA취류분석,증실해서렬시28S rRNA,동시환발현모두귀산적계통진화관계비교리산.차외,재저일신28S rRNA여TMV적항성단백기인y3지간발현유량개동원구단유가능시PCR확증y3기인시출현비목적조대적원인.재저량개동원구단중,기일구단여극륭y3기인시소용적PCR인물지일유교고적상사성,령일구단야시일반PCR인물적유사물.본연구중신28S rRNA서렬적획득시PCR확증중출현비목적조대적신례,해서렬적발현급취류분석적결과유조우진균기인조학연구급진균생물분자분류계통적건립.
In this study, we have found a new 28S rRNA sequence with 906 bp in length (GenBank accession No. GU568178) which isolated from mycelia ofbasidiomycete shaggy mane (Coprinus comatus), and the sequence was an untargeted band obtained unexpectedly when we previously cloned TMV-inhibitor y3 gene from this shaggy mane. The acquired sequence was used to search homologous sequences by using BLAST algorithm in NCBI, and it was performed to cluster with its homologues by using the Clustal w and MEGA methods. The results confirmed that the sequence was a new 28S rRNA of ribosomal large-subunits, and the phylogenic relationship ofCoprinus comatus was found to be dispersed from one anothers. Moreover, two homologous counterparts between the sequences of new 28S rRNA and the TMV-inhibitor gene y3 were found possibly to be the cause of formation of the untargeted band in PCR amplification of y3 gene. Within these two homologous counterparts, one counterpart had the sequence similar to a PCR primer for y3 gene, and the other one was a primer-like fragment. In this research, obtaining of the new se-quence of 28S rRNA was an updated example of the formation ofuntargeted bands during PCR amplification. Then the finding and clustering analysis results of 28S rRNA would be helpful for researches in fungus genomics and the establishment of theirs molecular classification system.