| 摘要: |
| 为筛选在鸡基因表达时稳定表达的内参基因，本试验以6、14、22及30周龄的地方品种固始鸡与6周龄的商业品种罗斯308肉鸡为试验动物，以B2M、28S、GAPDH、β-actin及HSP70为候选内参基因，利用qRT-PCR对这5个候选内参基因的相对表达量进行测定，通过独立评价软件geNorm、NormFinder、SPSS和Ct值分析法筛选出在不同组织、不同发育时期和不同品种间稳定表达的最佳内参基因。结果表明:在鸡不同组织中，28S为最佳内参基因；在鸡不同发育时期中，GAPDH为最佳内参基因；在不同品种鸡中最佳内参基因组合为GAPDH和28S。综上所述，在以地方品种固始鸡为例的不同组织，不同时期间最佳内参基因分别为28S和GAPDH；以地方品种固始鸡和商业品种罗斯308肉鸡为例的不同品种间最佳内参基因为GAPDH+28S基因组合。综上，本研究筛选出了鸡不同试验条件下的最佳内参基因，为规范鸡基因定量表达分析中内参基因的使用提供参考依据。 |
| 关键词: 鸡 内参基因 geNorm NormFinder |
| DOI：10.11841/j.issn.1007-4333.2021.04.07 |
| 投稿时间：2020-08-10 |
| 基金项目:国家自然科学基金项目(No.31902144)；河南省重点研发项目(192102110070)；2020年度河南省高等学校重点科研项目(20B23007) |
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| Screening optimum reference gene for gene expression in chicken by quantitative real-time PCR |
|
LI Yiyi,ZHANG Yanhua,WU Junfeng,ZHANG Chenxi,KANG Xiangtao,LI Wenting
|
| (College of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450000, China) |
| Abstract: |
| In order to screen the internal reference genes stably expressed in chicken, the local Gushi chicken of 6, 14, 22, and 30-week-old and the commercial variety Ross 308 of 6-week-old were used as experimental animals. Using B2M, 28S, GAPDH, β-actin and HSP70 as candidate reference genes, the relative expression levels of these five candidate reference genes were determined by qRT-PCR. by the three softwares(geNorm, NormFinder and SPSS)and Ct value analysis. The best internal reference genes were screened out in different tissues, different development stages and different varieties. The results showed that 28s was the best internal reference gene in different tissues of chicken; GAPDH was the best internal reference gene in different development stages; GAPDH and 28s were the best combination of internal reference genes in different chicken varieties. In summary, the best internal reference genes under experimental conditions were identified in this study, which provided a reference for standardizing the use of internal reference genes in chicken genes for quantitative expression analysis. |
| Key words: chicken internal reference gene geNorm NormFinder |
