Arabidopsis wild-type vs. vip3

Comparison of wild-type and vip3. The Arabidopsis thaliana VERNALIZATION INDEPENDENCE (VIP) gene class has multiple functions in development, including repression of flowering through activation of MADS box gene FLC. Among VIP genes, VIP genes encode yeast Paf1 complex homolog which is required for histone modification, transcriptional elongation. To identify genes regulated by VIP3, we performed Affymetrix Arabidopsis gene chip analysis and compared gene transcription profiles from wild-type and vip3 mutant plants. We found more than 200 genes misregulated in vip3 mutant plants compring to wild-type plant. Keywords: Comparison of wild -type and vip3 mutants Plants were grown on soil. Total aerial tissues were harvestedfrom 14 day old seedling. Each of two replicates for each genotypes was composed of three independently derived samples. Each sample included more than 100 plants. Total RNA was isolated using Qiagen RNAeasy columns (Qiagen, Valencia, CA). Synthesis of cDNA employed the SuperScript double-stranded cDNA sysnthesis kit (Affymetrix). Synthesis of biotinylated cDNA utilized the BioArray high yield RNA transcript labeling kit (Affymetrix). Hybridization and scanning of microarrays was performed at the Research Techonology Support Facility at Michigan State University. Microarray data were analyzed using the statistical algorithms within the Affymetrix Microarray Suite (MAS) 5.0 software.

野生型与vip3突变体的比较分析
拟南芥春化不依赖（VERNALIZATION INDEPENDENCE, VIP）基因家族在植物发育过程中具有多种功能，包括通过激活MADS盒基因FLC抑制开花。在VIP家族基因中，VIP基因编码酵母Paf1复合物的同源蛋白，该复合物对于组蛋白修饰与转录延伸不可或缺。
为鉴定受VIP3调控的基因，我们开展了Affymetrix拟南芥基因芯片分析，并对比了野生型与vip3突变体植株的基因转录谱。我们发现，相较于野生型植株，vip3突变体植株中有超过200个基因的表达失调。
关键词：野生型与vip3突变体的比较
供试植株均种植于土壤中，于幼苗生长14天时收获其全部地上组织。每种基因型设置2次生物学重复，每次重复由3份独立制备的样本组成，每份样本包含超过100株植株。总RNA提取采用Qiagen RNAeasy柱式提取试剂盒（Qiagen，美国加利福尼亚州瓦伦西亚）。cDNA合成采用SuperScript双链cDNA合成试剂盒（Affymetrix）。生物素标记cDNA的合成采用BioArray高产率RNA转录物标记试剂盒（Affymetrix）。芯片杂交与扫描工作均在密歇根州立大学研究技术支持中心完成。芯片数据分析采用Affymetrix Microarray Suite（MAS）5.0软件内置的统计算法。