Gene expression C. elegans age-1 mutants with neuron- or intestine-restricted age-1 activity

The goal of this experiment was to identify transcripts regulated non-autonomously by age-1 activity in neurons or intestine. Comparisons of gene expression were made between age-1(mg44) null adults and CY251 (age-1(mg44); bvIs2) and CY262 (age-1(mg44); bvIs1); all were relative to wildtype (N2). The integrated transgenes, bvIs2 and bvIs1, carry the age-1 cDNA expressed from the neuronal ric-19 promoter (bvIs2) or intestinal ges-1 promoter (bvIs1). Transcripts upregulated in age-1(mg44) adults compared to wildtype (&gt;=1.5 fold; p&lt;0.05) or downregulated (&lt;0.6 fold; p&lt;0.05) were considered to be age-1 target genes. Rescue in CY251 or CY262 was determined by change in the transgenic background versus age-1(mg44) (all relative to N2 wildtype). Non-autonomous rescue of a target gene was usually determined by transcript levels that were at least 50% rescued compared to wildtype.本实验旨在探究神经元或肠道中由age-1活性介导的非自主性调控转录本。实验以野生型（N2）为参照，对age-1(mg44)纯合缺失成虫、CY251（age-1(mg44); bvIs2）及CY262（age-1(mg44); bvIs1）的基因表达水平进行了比较分析。两处整合转基因位点bvIs2与bvIs1，分别搭载了由神经元特异性ric-19启动子、肠道特异性ges-1启动子驱动表达的age-1互补DNA（cDNA）。以转录本表达量较野生型上调≥1.5倍且显著性p<0.05，或下调≤0.6倍且显著性p<0.05作为age-1靶基因的判定依据。CY251与CY262的基因挽救效果，通过其转基因背景相较于age-1(mg44)的表达变化进行评估（所有比较均以野生型N2为参照）。靶基因的非自主性挽救通常以其转录本水平较age-1(mg44)至少恢复至野生型水平的50%作为判定标准。