βactin基因在荒漠昆虫光滑鳖甲和小胸鳖甲中表达稳定性研究.docVIP

βactin基因在荒漠昆虫光滑鳖甲和小胸鳖甲中表达稳定性研究 　　摘 要：为检测生活习性截然相反的两种荒漠昆虫光滑鳖甲和小胸鳖甲的β-actin基因在不同温度下的表达是否稳定，对不同温度处理和不同季节采集的昆虫提取总RNA反转录合成cDNA后，进行荧光实时定量PCR检测。以Ct值作为β-actin基因表达水平的测定值。通过方差分析显示：光滑鳖甲的β-actin基因在不同季节无显著差异，而小胸鳖甲则表现出β-actin基因表达的季节性差异；在不同温度处理条件下，光滑鳖甲β-actin基因表达比小胸鳖甲的要稳定。研究认为β-actin可作为研究光滑鳖甲不同温度下基因表达的内参基因，而β-actin则不适合用做研究小胸鳖甲在不同温度下基因表达的内参基因。 　　关键词：β-actin； 内参基因；光滑鳖甲；小胸鳖甲；实时定量PCR 　　中图分类号：S188 文献标识码：A DOI编码：10.3969/j.issn.1006-6500.2014.02.001 　　Expression Stability of β-actin Gene in the Desert Insects Microdera punctipennis and Anatolica polita 　　MENG Shan-shan，MA Wen-jing，LIU Xiao-ning，MA Ji 　　（Xinjiang Key Laboratory of Biological Resources and Genetic Engineering， College of Life Science and Technology， Xinjiang University， Urumqi，Xinjiang 830046， China） 　　Abstract： In order to measuring the expression stability of the β-actin gene at different temperatures in two desert insects， Microdera punctipennis and Anatolica polita， which have quite contrary living habit，total RNA from the insects collected in different seasons， or treated at different temperatures was extracted and reverse transcribed to synthesize cDNAs. The expression of the β-actin genes from these two species was measured by Ct values which were obtained by real-time quantitative PCR. The results showed that there were no significant changes in the expression of β-actin gene in Anatolica polita over seasons， while in Microdera punctipennis the expression of β-actin gene changed significantly. When the beetles were treated at different temperatures， the expression of β-actin gene in Anatolica polita was more stable than that in Microdera punctipennis. β-actin gene in Anatolica polita could be used as a reference gene for study gene expression at different temperatures， while it is not suitable in Microdera punctipennis for study gene expression at different temperatures. 　　Key words： β-actin； reference gene； Microdera punctipennis； Anatolica polita； real-time PCR 　　β-actin是肌细胞骨架微丝的主要成分，具有收缩功能，参与胞质分裂、形态维持、生