江西农业大学学报2012，Vol.34Issue(3)：580-584,5.
建鲤内参基因EF-1α的实时荧光定量PCR方法的建立
Establishment of a Real Time PCR Assay for Cyprinus carpio var.jian EF-1a As a Reference Gene
摘要
Abstract
Eukaryotic elongation factor la (EF-lα) plays an important role in translation and its sequence is highly conservative as a housekeeping gene in real time PCR. The partial cDNA encoding EF-la in Cyprinus carpio var. jian was isolated using RT-PCR. The sequence of cDNA was 425 bp in length encoding 140 amino acids residues, and the homology was about 98% between C. carpio var. jian and other fish using the Blast program. The DNA sequence of C. carpio var. jian EF-1 a consisting of 506 bp was also cloned. Comparing the partial cDNA to its genomic sequence revealed that C. carpio var. jian EF-la gene consisted of a phase 0 intron. A pair of real time PCR primers cross intron was designed according to the intron sequence of EF-1 a and a real time PCR method by SYBR Green I was established. The standard curve was established with liver cDNA as the standard template and the melting analysis was also carried out. The results showed that the real time PCR method for EF-1 a had the advantages of high specificity, good correlation coefficients and wide linear range, which supplied useful information for studying function gene expression.关键词
建鲤/内参基因/EF-1α/实时荧光定量PCRKey words
Cyprinus carpio vai. jian, housekeeping gene, EF-la, real time PCR分类
农业科技引用本文复制引用
唐永凯,俞菊华,徐跑,李建林,李红霞,董在杰,夏正龙..建鲤内参基因EF-1α的实时荧光定量PCR方法的建立[J].江西农业大学学报,2012,34(3):580-584,5.基金项目
农业部公益性行业专项(200903045)、基本科研业务费专项资金(2011JBFA13)和现代农业产业技术体系(nycytx-49) （200903045）
