Evaluation of Several Endogenous Reference Genes in Streptomyces diastatochromogenes 1628 and Its Drug Resistance Mutants

doi:10.16409/j.cnki.2095-039x.2016.04.014

Abstract

Abstract: Metabolites of Streptomyces diastatochromogenes 1628 have strong inhibition on various plant pathogenic fungi. To screen out the most stably expressed endogenous reference genes in different drug resistance overproduction mutants of S. diastatochromogenes 1628, six candidate reference genes were selected for real-time quantitative PCR, and two kinds of software were adopted to determine the stability of candidate reference genes in different drug resistance strains. The results revealed that the optimum reference gene analyzed by different softwares was slightly different. Analysis of geNorm software showed the gyrB, gyrB, thyA, rpoA were the most stably expressed reference gene in wild-type, streptomycin-resistance, rifampin-resistance and paromomycin- resistance strain, respectively. However, rpoA was the most stable gene in tested strains analyzed by NormFinder. Comprehensively, the mean rank based on stability banking analyzed by these two kinds of software showed that rpoA was the most suitable reference gene in all tested strains. By detecting the expression level of toyG gene, it was found that rpoA as internal control gene is more reasonable.

Key words: Streptomyces diastatochromogenes, qPCR, reference gene

CLC Number:

S476

TANG Gu, YAO Jiayi, SHENTU Xuping, YU Xiaoping. Evaluation of Several Endogenous Reference Genes in Streptomyces diastatochromogenes 1628 and Its Drug Resistance Mutants[J]. journal1, 2016, 32(4): 518-523.

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