鹿茸组织中内参基因的筛选和验证

doi:10.16431/j.cnki.1671-7236.2015.04.017

›› 2015, Vol. 42 ›› Issue (4): 883-889.doi: 10.16431/j.cnki.1671-7236.2015.04.017

鹿茸组织中内参基因的筛选和验证

张然然¹, 刘华淼¹, 邢秀梅¹, 胡大勇²

1. 中国农业科学院特产研究所, 特种经济动物分子生物重点实验室, 长春 130112;
2. 白城市动物卫生监督所, 白城 137000

收稿日期:2014-09-19 出版日期:2015-04-20 发布日期:2015-05-05
通讯作者: 邢秀梅 E-mail:xingxiumei2004@126.com
作者简介:张然然(1990-),女,河北深州人,硕士生,研究方向:鹿茸蛋白质组学,E-mail:heavenranran@163.com
基金资助:
特种动物遗传资源创新团队(CAAS-ASTIP-201X-ISAPS);吉林省科技攻关计划(20140204058NY)

Selection and Validation of Reference Genes in Velvet Antlers Tissues

ZHANG Ran-ran¹, LIU Hua-miao¹, XING Xiu-mei¹, HU Da-yong²

1. State Key Laboratory of Special Economic Animal Molecular Biology, Institute of Special Wild Economic Animal and Plant Science, Chinese Academy of Agricultural Sciences, Changchun 130112, China;
2. Jilin Province Baicheng City Animal Health Supervision, Baicheng 137000, China

Received:2014-09-19 Online:2015-04-20 Published:2015-05-05

摘要/Abstract

摘要： 为筛选在不同生长时期鹿茸组织中稳定表达的内参基因,试验以不同生长时期(分别为脱盘后10、20、40和60 d)的鹿茸组织为材料,采用实时荧光定量PCR(qRT-PCR)方法分析甘油醛-3-磷酸脱氢酶(GAPDH)、β2-微球蛋白(B2M)、还原型辅酶Ⅰ(NADH)、60S 核糖体蛋白L40(RPL40)、谷胱甘肽还原酶7(GPx)和β肌动蛋白(ACTB)6个看家基因的表达情况,并运用 geNorm和NormFinder 两个程序综合分析6个看家基因的表达稳定性.结果显示,GAPDH、ACTB、RPL40表达稳定性较好,可用作鹿茸基因表达研究的内参基因,而NADH和GPx的稳定性最差,不适合作内参基因.通过对鹿茸生长相关基因(ANXA5、HSP27、PRD2、CRABP1、LGALS1)表达分析,进一步验证了上述结果,并且发现这5种基因均在脱盘后10 d的鹿茸组织中高表达.该研究结果为鹿茸快速生长及骨化相关基因的研究奠定了一定基础.

关键词: qRT-PCR; 鹿茸; 内参基因; geNorm; NormFinder

Abstract: The aim of this study was to identify the most stable gene to be used as reference genes for qRT-PCR analysis in Sika deer antler tissues. The expression patterns of 6 housekeeping genes were analyzed, including glyceraldehyde-3-phosphate dehydrogenase (GAPDH), beta-2-microglobulin (B2M), NADH dehydrogenase (NADH), 60S ribosomal protein L40 (RPL40), glutathione peroxidase 7 (GPx) and beta actin (ACTB), in Sika deer antler at different stages (10, 20, 40 and 60 d). The stability of housekeeping gene expression was analyzed with use of 2 software packages, including geNorm and NormFinder. The results showed that GAPDH, ACTB, RPL40 were suitable reference genes for efficient normalization of qRT-PCR data, whereas NADH and GPx were not suitable for analysis of Sika deer antler. These findings were confirmed by comparative profiling of 5 antler genes associated with antler rapid growth (ANXA5, HSP27, PRD2, CRABP1 and LGALS1), while it was observed that the 5 genes were significanly expressed in antler aged 10 d. These results will provide a necessary basis for the further research on rapid growth of antler and ossification genes.

Key words: qRT-PCR; velvet antlers; reference genes; geNorm; NormFinder

中图分类号:

S825

张然然, 刘华淼, 邢秀梅, 胡大勇. 鹿茸组织中内参基因的筛选和验证[J]. , 2015, 42(4): 883-889.

ZHANG Ran-ran, LIU Hua-miao, XING Xiu-mei, HU Da-yong. Selection and Validation of Reference Genes in Velvet Antlers Tissues[J]. , 2015, 42(4): 883-889.

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鹿茸组织中内参基因的筛选和验证

Selection and Validation of Reference Genes in Velvet Antlers Tissues

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