浅黄根须腹菌γ-actin基因的克隆及表达分析

doi:10.11707/j.1001-7488.20141011

Abstract

Abstract:

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.

Key words: Rhizopogon luteolus, actin gene, cDNA, gene cloning, gene expression

CLC Number:

Q933

Zheng Rong, Wang Jugang, Tai Lihua, Bai Shulan, Niu Yanfang. Cloning and Expression Analysis of γ-actin Gene from Rhizopogon luteolus[J]. Scientia Silvae Sinicae, 2014, 50(10): 80-85.

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Cloning and Expression Analysis of γ-actin Gene from Rhizopogon luteolus

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