کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5906263 | 1159967 | 2013 | 10 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Evaluation of reference genes for quantitative real-time RT-PCR analysis of gene expression in Nile tilapia (Oreochromis niloticus)
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کلمات کلیدی
LTACathepsin ZUBCECTSZb2mEF1aPBSGADPH18S rRNACTSDRT-qPCRLPS18S ribosomal RNA - RNA ریبوزوم 18Scycle threshold - آستانه چرخهubiquitin-conjugating enzyme - آنزیم کنگوتیک ubiquitinlipoteichoic acid - اسید لیپتویکوئیکbeta actin - بتا آکتینBeta-2-microglobulin - بتا-2-میکروگلوبولینTuba - توباTilapia - تیلاپیاelongation factor 1 alpha - ضریب اصطکاک 1 آلفاCorrelation coefficients - ضریب همبستگیActb - عملlipopolysaccharide - لیپوپلی ساکاریدPhosphate-buffered saline - محلول نمک فسفات با خاصیت بافریReference genes - ژنهای مرجعcathepsin D - کاتهپسین DPCR efficiency - کارایی PCRglyceraldehyde-3-phosphate dehydrogenase - گلیسرالیدید-3-فسفات دهیدروژناز
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
ژنتیک
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Quantitative real-time reverse-transcriptase polymerase chain reaction (RT-qPCR) has been used frequently to study gene expression related to fish immunology. In such studies, a stable reference gene should be selected to correct the expression of the target gene. In this study, seven candidate reference genes (glyceraldehyde-3-phosphate dehydrogenase (GADPH), ubiquitin-conjugating enzyme (UBCE), 18S ribosomal RNA (18S rRNA), beta-2-microglobulin (B2M), elongation factor 1 alpha (EF1A), tubulin alpha chain-like (TUBA) and beta actin (ACTB)), were selected to analyze their stability and normalization in seven tissues (liver, spleen, kidney, brain, heart, muscle and intestine) of Nile tilapia (Oreochromis niloticus) challenged with Streptococcus agalactiae or Streptococcus iniae, respectively. The results showed that all the candidate reference genes exhibited tissue-dependent transcriptional variations. With PBS injection as a control, UBCE was the most stable and suitable single reference gene in the intestine, liver, brain, kidney, and spleen after S. iniae infection, and in the liver, kidney, and spleen after S. agalactiae infection. EF1A was the most suitable in heart and muscle after S. iniae or S. agalactiae infection. GADPH was the most suitable gene in intestine and brain after S. agalactiae infection. In normal conditions, UBCE and 18S rRNA were the most stably expressed genes across the various tissues. These results showed that for RT-qPCR analysis of tilapia, selecting two or more reference genes may be more suitable for cross-tissue analysis of gene expression.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Gene - Volume 527, Issue 1, 15 September 2013, Pages 183-192
Journal: Gene - Volume 527, Issue 1, 15 September 2013, Pages 183-192
نویسندگان
Chang Geng Yang, Xian Li Wang, Juan Tian, Wei Liu, Fan Wu, Ming Jiang, Hua Wen,