蛋白激酶Cla4对黄曲霉生长发育、毒素合成和致病力的影响

秦岭,李晓雯,李丁,赵家儒,汪世华,袁军

Protein kinase Cla4 regulates morphology development, aflatoxin biosynthesis and pathogenicity of Aspergillus flavus

QIN Ling,LI Xiao-Wen,LI Ding,ZHAO Jia-Ru,WANG Shi-Hua,YUAN Jun

图2 黄曲霉Aflcla4基因缺失菌株的构建 A：Aflcla4基因敲除原理示意图;B：PCR扩增ΔAflcla4的ORF,AP和BP片段（引物为P9/P10,P2/P907和P919/P7）;C：Aflcla4 敲除菌株的反转录PCR验证,actin基因为对照;D：ΔAflcla4和WT菌株的qRT-PCR表达量分析,柱状图代表Aflcla4基因在菌株中的相对表达量. 2-ΔΔCt法用于计算目标基因的表达量,β-actin基因作为内参. ND：未检测到

Fig. 2 Generation of cla4 deletion mutant in Aspergillus flavus. A: Schematic illustration for Aflcla4 disruption; B: The results of the PCR analysis of ΔAflcla4, ORF (open reading frame) AP and BP fragments were amplified with a couple of primers P9/P10, P2/P907 and P919/P7 respectively. C: Verification of Aflcla4 deletion strain with reverse transcription-PCR. Actin gene was used as an endogenous control. D: qRT-PCR analysis of the expression level of Aflcla4 gene in ΔAflcla4 mutant and WT (wild-type) strains. The histogram (D) indicates relative expression level of Aflcla4 gene in strains. The 2-ΔΔCt method was used to evaluate target gene expression levels, which were relative to the expression level of the β-actin reference gene. ND: Not detected.