摘要
采用双标准曲线相对实时荧光定量PCR法,分别以23℃常温对照组和6℃低温待测组的黑龙江鲤脑组织cDNA为模板,以18S rRNA为内参基因,检测26个候选基因的相对表达量。实验数据经显著性分析发现,有5个候选基因在低温条件下表达量显著上升(P<0.01),与对照组相比它们的表达量分别上升了2.11倍、13.9倍、2.52倍、7.38倍和1.83倍,基因功能比对结果表明其编码蛋白产物分别是脂肪酸链延伸蛋白、酰基辅酶A脱氢酶、转录起始因子IIB、肌醇-1-磷酸合成酶、血脑屏障HT7抗原;有7个候选基因在低温下表达量分别下降了21.8%、25.9%、16.6%、23.7%、15.8%、16.3%、42.5%,但对照组和待测组差异不显著(P>0.05),基因功能比对发现它们主要参与抑制糖酵解,促进细胞凋亡和干扰神经系统的重塑活动。上述低温下表达量显著上升的5个冷诱导候选基因的获得为今后进行不耐低温鱼类的基因工程育种提供了基因元件。
In this study,26 candidate genes were quantified and normalized in the brain cDNA of common carp（Cyprinus carpio） at 23°C and 6°C using double-standard curve method of real-time quantitative PCR.The results showed that five candidates up-regulated in the samples at 6°C（P0.01） and quantified 2.11,13.9,2.52,7.38,and 1.83 times more than in the samples at 23°C,respectively.Gene function searching indicated that the protein products of these five candidates were elongation of very long chain fatty acids protein,Acyl-CoA desaturase,Transcription initiation factor IIB,Myo-inositol-1-phosphate synthase,and Blood-brain barrier HT7 antigen individually.Moreover,seven down-regulated candidates were also identified in the same samples at 6°C（P0.05）,and their expression levels were decreased by 21.8%,25.9%,16.6%,23.7%,15.8%,16.3%,and 42.5%,respectively,in comparison with the samples at 23°C.These seven down-regulated can-didates mainly participated in the inhibition of glycolysis,improvement of cell apoptosis,and intervention of synapse re-modeling based on the results of function searching.The five cold-induced genes identified in this study will be used as important elements for fish with cold sensitive through transgenic technology in future.
出处
《遗传》
CAS
CSCD
北大核心
2011年第3期262-269,共8页
Hereditas（Beijing)
基金
国家重点基础研究发展计划"973计划"项目(编号:2010CB126305)
农业部水产生物技术重点开放实验室项目(编号:2009NYBZS-05)
中央级公益性科研院所基本科研业务费专项资金项目(编号:2009HSYZX-SJ-05)资助
关键词
鲤
脑组织cDNA
低温
候选基因
实时荧光定量PCR
双标准曲线法
Cyprinus carpio
brain cDNA
low temperature
candidate genes
real-time quantitative PCR
double-standard curves
