中国农业科学院农业信息研究所 中国农学会
Agricultural Information Institute of CAAS China Association of Agricultural Science Societies
En
Ch
中国农业科学院农业信息研究所 中国农学会 主办

AI检索

订阅

返回顶部
当前位置: 首页 > 文章 > 米蛾β-actin基因cDNA片段的克隆及表达量检测 沈阳农业大学学报 2016,47 (1) 24-28
Position: Home > Articles > Molecular Cloning and Expression Levels of β-actin Gene in Rice Moth [Corcyra cephalonica(Stainton)] Journal of Shenyang Agricultural University 2016,47 (1) 24-28

米蛾β-actin基因cDNA片段的克隆及表达量检测

作  者：
丛斌;张明珠;胡志凤;段立佳;王晓静;张统书;董辉
单  位：
沈阳农业大学植物保护学院
关键词：
米蛾;β-actin基因;反转录PCR;实时荧光定量PCR
摘  要：
为克隆米蛾[Corcyra cephalonica(Stainton)]β-actin基因cDNA片段,建立米蛾β-actin基因实时荧光定量PCR(qRT-PCR)方法,并评估其在实时荧光定量研究中作为内参基因的可靠性,利用反转录PCR(RT-PCR)技术克隆获得米蛾β-actin基因cDNA片段,采用SYBR GreenⅠ染料法建立qRT-PCR方法,检测在米蛾卵、幼虫、蛹和成虫4个虫态中β-actin基因的表达量。获得了长为822bp的米蛾β-actin基因片段(Gen Bank accession:KJ599569),编码273个氨基酸残基;荧光定量标准曲线的Ct值检测范围为13~28,扩增效率为90.1%,相关系数R~2=0.992,溶解曲线分析结果显示产物为特异的单峰,其Tm值为(84±0.5)℃,符合定量要求;实时荧光定量结果表明,β-actin基因在米蛾不同发育历期中的表达水平无显著性差异(p>0.05)。成功建立了米蛾β-actin基因qRT-PCR方法 ,β-actin基因可以作为米蛾基因表达定量研究中的可靠内参基因。研究结果为β-actin作内参基因进行米蛾功能基因表达差异研究奠定了分子基础。
译  名：
Molecular Cloning and Expression Levels of β-actin Gene in Rice Moth [Corcyra cephalonica(Stainton)]
作  者：
CONG Bin;ZHANG Ming-zhu;HU Zhi-feng;DUAN Li-jia;WANG Xiao-jing;ZHANG Tong-shu;DONG Hui;College of Plant Protection, Shenyang Agricultural University;
关键词：
Corcyra cephalonica;;β-actin gene;;reverse transcription PCR;;real-time fluorescence quantitative PCR
摘  要：
We cloned the cDNA of β-actin gene fragment from Corcyra cephalonica(Stainton) and evaluated the reliability of this gene as a reference gene in the Real-time fluorescent quantitative PCR(qRT-PCR) researchs. β-actin gene fragment was cloned by using reverse transcription PCR(RT-PCR) technique, qRT-PCR was performed using SYBR Green I to detect the expression of β-actin in four different stages of C. cephalonica. The results showed that the β-actin cDNA gene fragment had 822 base pairs in length(Gen Bank accession: KJ599569) and encoded 273 amino acid residues. The Ct value range of fluorescence quantitative standard was about 13-28, the amplification efficiency was 90.8%, R~2=0.993, Tm value was(84±0.5)℃. These numbers conform to the requirements of the quantitative. The qRT-PCR results showed that expression level was close in different stages of C.cephalonica. The method of qRT-PCR of β-actin from C. cephalonica was established successfully, β-actin gene could be used as a reliable reference gene in the quantitative analysis of gene expression in C. cephalonica. The research of β-actin laid the molecular foundation for the further research on the different expression of function genes in C. cephalonica.

相似文章

导出引用
计量
文章访问数: 10
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊

友情链接Friendly link
农业信息研究所Agricultural information institute 农业智能知识服务平台Agricultural intelligent knowledge service platform 农业学术服务平台Agricultural Knowledge Service System 中国知网CNKI 维普网VIP 万方WANFANG DATA
联系我们Contact us

电话Tell：010-82109652-808

邮箱E-mail：agrijournal@caas.cn

服务Service
关于我们About us 意见建议Opinions and suggest
版权说明Copyright
单位：中国农业科学院农业信息研究所 中国农学会 © 2017中国农业科学院农业信息研究所 ICP备案号：京ICP备09089781号-46