The study of KpsF gene which contains PxIxIT motif in Aspergillus fumigatus

LI Jia-juan; LI Wen; MA Yan

doi:10.3969/j.issn.1673-3827.2017.01.003

Abstract：Objective To construct the KpsF gene defect strains △KpsF and to explore the interaction between KpsF gene and calcineurin.Methods Amplificated the upstream and downstream of KpsF about 1.0 kb DNA fragments,the pyrG of plasmid pJW 24 was used as a biomarker when constructting the knockout plasmid.Recombinant plasmid was transformed into A.fumigatus Ku80 pyrG and got △KpsF.To observer the radial and pbenotypic growth of Ku80 and △KpsF,Real-time PCR was used to determine the relative expression of calcineurin catalytic subunit (CnaA) of Ku80 (control group)and △KpsF (experimental group) in GMM liquid medium containing different concentration of CaCl2.Results Compared with Ku80,the radial growth of △KpsF had no obvious change (P＞0.05),and the phenotypic growth had no obvious difference.In GMM Liquid medium containing 10 mmol/L CaCl2,the relative expression of CnaA of △KpsF strain was significantly higher than Ku80 (P＜0.01).However,the relative expression of CnaA did not obviously changed (P＞0.05) in GMM Liquid medium containing 100 mmol/L CaCl2.Conclusions The KpsF had no effect in radial and phenotypic growth of A.fumigatus.Under the condition of different concentrations of Ca2+,the relative expression levels of CnaA gene were different between △KpsF and Ku80,suggesting that the KpsF might be involved in the feedback regulation of calcineurin under the condition of low concentration of CaCl2.