mRNA差异显示技术已经广泛应用于研究植物、动物和微生物在各种环境条件下基因的差异表达研究.研究以15℃和30℃培养的深黄被孢霉M6-22菌体为材料,利用mRNA差异显示技术研究两种培养条件下基因的表达差异.经实时荧光定量PCR验证,共获得7条差异片段,相似性搜索结果表明它们是6-磷酸葡萄糖异构酶、单糖核苷酸转运蛋白、Ras1鸟苷酸转移因子、依赖于NAD的苹果酸脱氢酶、Δ^12-脂肪酸脱氢酶、CLK4关联的丝氨酸/精氨酸丰富蛋白和假定蛋白,涉及糖酵解、蛋白质修饰、信号传导、脂肪酸合成和mRNA加工等生命过程,表明深黄被孢霉M6-22低温适应性是多种途径协同调控的结果.
The mRNA differential display PCR is widely used to investigate the differential gene expression in response to different environments in plants,animals and microorganisms.In this study,the technique was used to analyze the differential gene expression in Mortierella isabellina strain M6-22 which was cultured at 15 ℃ and30 ℃,respectively.The results showed that 7 fragments whose mRNA levels were increased by 2 times or more at15 ℃ were obtained from 21 candidate fragments after verification by real-time quantitative PCR. They showed high amino acid sequence similarity to glucose-6-phosphate isomerase,sugar-nucleotide transporter,ras1 guanine nucleotide exchange factor,NAD dependent malate dehydrogenase,Δ^12-fatty acid desaturase and CLK4-associating serine / arginine rich protein,being potentially involved in the life processes of glycolysis,protein modification,signal transduction,fatty acid biosynthesis and mRNA processing,et al.These results indicate that the adaptation of M. isabellina M6-22 to low temperature is a coordinate regulation of multiple pathways.