CAJ | 학술논문

以油松的优良外生菌根真菌即浅黄根须腹菌为对象，用简并 PCR 法和 RACE 技术分离其γ－肌动蛋白基因( Rl-act)的全长 cDNA序列。该全长序列为1339 bp，包含一个1128 bp 的开放阅读框( ORF)，编码375个氨基酸，5'端非翻译区(5'UTR)95 bp，3'UTR长度116 bp。Port Param 软件在线分析结果表明，该cDNA所编码的蛋白质理论等电点为5.01，相对分子质量为94.929 kD，具有真菌γ-actin 基因3个保守特征序列。Blast同源性检索结果表明，Rl-act序列与12种担子菌 actin 序列同源性均在97%以上，与同为外生菌根真菌的双色蜡蘑actin的亲缘关系最近。Rl-act基因在不同碳源及磷水平培养条件下表达量基本一致，验证了该基因作为分子内标的可靠性。
이유송적우량외생균근진균즉천황근수복균위대상，용간병 PCR 법화 RACE 기술분리기γ－기동단백기인( Rl-act)적전장 cDNA서렬。해전장서렬위1339 bp，포함일개1128 bp 적개방열독광( ORF)，편마375개안기산，5'단비번역구(5'UTR)95 bp，3'UTR장도116 bp。Port Param 연건재선분석결과표명，해cDNA소편마적단백질이론등전점위5.01，상대분자질량위94.929 kD，구유진균γ-actin 기인3개보수특정서렬。Blast동원성검색결과표명，Rl-act서렬여12충담자균 actin 서렬동원성균재97%이상，여동위외생균근진균적쌍색사마actin적친연관계최근。Rl-act기인재불동탄원급린수평배양조건하표체량기본일치，험증료해기인작위분자내표적가고성。
In this paper,a superior ectomycorrhizal fungus,Rhizopogon luteolus of Pinus tabulaeformis’was used as the research object. The fungus γ-actin gene( Rl-act) was isolated by using homology based method and RACE technique. The results showed that the full-length sequences of Rl-act cDNA 1 339 bp which was consisted of a 1 128 bp opening reading frame (ORF),encoding a protein of 375 amino acids,a 5'-UTR with 95 bp and a 3'-UTR with 116 bp. The online analysis of Port Param software displayed that the putative amino acids had an isoelectric point of 5. 01 ,a molecular weight of 94. 929 kD,and 3 highly conserved regions of γ-actin gene of fungi. Homology search indicated that the homogeneity was more than 97% between Rl-act cDNA and 12 basidiomycetes actin sequence. Phylogenetic tree indicated that it had closer relationship with Laccaria bicolor,an ectomycorrhizal fungus. In different culture conditions of carbon and phosphorus level,the quantity expression of Rl-act was almost same,which validated the reliability of Rl-act as reference gene in quantitative analysis of mRNA expression. The research provided an internal standard for studying other stress resistance genes’ expression and regulation in symbiosis of R. luteolus and P. tabulaeformis,and gave a theoretical foundation for exploring the molecular mechanism in stress resistance.