Acetolactate Synthase Overexpression in Mesosulfuron-Methyl-Resistant Shortawn Foxtail (Alopecurus aequalis Sobol.): Reference Gene Selection and Herbicide Target Gene Expression Analysis

2018 ◽  
Vol 66 (37) ◽  
pp. 9624-9634 ◽  
Author(s):  
Ning Zhao ◽  
Yanyan Yan ◽  
Hengzhi Wang ◽  
Shuang Bai ◽  
Qian Wang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Reference Gene ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Target Gene ◽  
Gene Selection ◽  
Acetolactate Synthase ◽  
Target Gene Expression ◽  
Reference Gene Selection

scholarly journals Reference Gene Selection for Gene Expression Analysis of Oocytes Collected from Dairy Cattle and Buffaloes during Winter and Summer

PLoS ONE ◽  
2014 ◽  
Vol 9 (3) ◽  
pp. e93287 ◽  
Author(s):  
Carolina Habermann Macabelli ◽  
Roberta Machado Ferreira ◽  
Lindsay Unno Gimenes ◽  
Nelcio Antonio Tonizza de Carvalho ◽  
Júlia Gleyci Soares ◽  
...  
Keyword(s):  
Gene Expression ◽  
Dairy Cattle ◽  
Reference Gene ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Gene Selection ◽  
Reference Gene Selection ◽  
Selection For

scholarly journals Reference Gene Selection for Normalizing Gene Expression in Ips Sexdentatus (Coleoptera: Curculionidae: Scolytinae) Under Different Experimental Conditions

2021 ◽  
Vol 12 ◽  
Author(s):  
Gothandapani Sellamuthu ◽  
Shan Amin ◽  
Jan Bílý ◽  
Jirí Synek ◽  
Roman Modlinger ◽  
...  
Keyword(s):  
Gene Expression ◽  
Reference Gene ◽  
Expression Analysis ◽  
Reference Genes ◽  
Gene Expression Analysis ◽  
Gene Selection ◽  
Experimental Conditions ◽  
Tissue Specific ◽  
Reference Gene Selection ◽  
Ips Sexdentatus

Ips sexdentatus (Coleoptera: Curculionidae: Scolytinae) is one of the most destructive and economically important forest pests. A better understanding of molecular mechanisms underlying its adaptation to toxic host compounds may unleash the potential for future management of this pest. Gene expression studies could be considered as one of the key experimental approaches for such purposes. A suitable reference gene selection is fundamental for quantitative gene expression analysis and functional genomics studies in I. sexdentatus. Twelve commonly used reference genes in Coleopterans were screened under different experimental conditions to obtain accurate and reliable normalization of gene expression data. The majority of the 12 reference genes showed a relatively stable expression pattern among developmental stages, tissue-specific, and sex-specific stages; however, some variabilities were observed during varied temperature incubation. Under developmental conditions, the Tubulin beta-1 chain (β-Tubulin) was the most stable reference gene, followed by translation elongation factor (eEF2) and ribosomal protein S3 (RPS3). In sex-specific conditions, RPS3, β-Tubulin, and eEF2 were the most stable reference genes. In contrast, different sets of genes were shown higher stability in terms of expression under tissue-specific conditions, i.e., RPS3 and eEF2 in head tissue, V-ATPase-A and eEF2 in the fat body, V-ATPase-A and eEF2 in the gut. Under varied temperatures, β-Tubulin and V-ATPase-A were most stable, whereas ubiquitin (UbiQ) and V-ATPase-A displayed the highest expression stability after Juvenile Hormone III treatment. The findings were validated further using real-time quantitative reverse transcription PCR (RT-qPCR)-based target gene expression analysis. Nevertheless, the present study delivers a catalog of reference genes under varied experimental conditions for the coleopteran forest pest I. sexdentatus and paves the way for future gene expression and functional genomic studies on this species.

scholarly journals Total RNA extraction from tissues for microRNA and target gene expression analysis: not all kits are created equal

2018 ◽  
Vol 18 (1) ◽  
Author(s):  
Rikki A. M. Brown ◽  
Michael R. Epis ◽  
Jessica L. Horsham ◽  
Tasnuva D. Kabir ◽  
Kirsty L. Richardson ◽  
...  
Keyword(s):  
Gene Expression ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Target Gene ◽  
Rna Extraction ◽  
Total Rna ◽  
Target Gene Expression ◽  
Total Rna Extraction

scholarly journals Selection of Reference Genes for Normalization of Real-Time PCR Data in Calliptamus italicus (Orthoptera: Acrididae) Under Different Temperature Conditions

2019 ◽  
Vol 19 (6) ◽  
Author(s):  
Hongxia Hu ◽  
Xiaofang Ye ◽  
Han Wang ◽  
Rong Ji
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Expression Analysis ◽  
Reference Genes ◽  
Gene Expression Analysis ◽  
Target Gene ◽  
Molecular Mechanisms ◽  
Expression Profiles ◽  
Quantitative Polymerase Chain Reaction ◽  
Target Gene Expression

Abstract Global warming has dominated worldwide climate change trends, and adaptability to high temperatures is the main factor underlying the spread of the pest Calliptamus italicus in Xinjiang Province, China. However, knowledge about the molecular mechanisms responsible for this adaptability and other related biological properties of C. italicus remain relatively unclear. Real-time quantitative polymerase chain reaction (RT-qPCR) is a key tool for gene expression analysis associated with various biological processes. Reference genes are necessary for normalizing gene expression levels across samples taken from specific experimental conditions. In this study, transcript level of five genes (GAPDH, 18S, TUB, ACT, and EF1α), commonly used as reference genes, were evaluated under nine different temperatures (27, 30, 33, 36, 39, 42, 45, 48, and 51°C) to assess their expression stability and further select the most suitable to be used on normalization of target gene expression data. Gene expression profiles were analyzed using geNorm, NormFinder, and BestKeeper software packages. The combined results demonstrated that the best-ranked reference genes for C. italicus are EF1α, GAPDH, and ACT under different thermal stress conditions. This is the first study that assesses gene expression analysis across a range of temperatures to select the most appropriate reference genes for RT-qPCR data normalization in C. italicus. These results should assist target gene expression analysis associated with heat stress in C. italicus.

scholarly journals Current Practices for Reference Gene Selection in RT-qPCR of Aspergillus: Outlook and Recommendations for the Future

Genes ◽  
2021 ◽  
Vol 12 (7) ◽  
pp. 960
Author(s):  
Meagan Archer ◽  
Jianping Xu
Keyword(s):  
Gene Expression ◽  
Reference Gene ◽  
Reference Genes ◽  
Gene Selection ◽  
Quantitative Polymerase Chain Reaction ◽  
Specific Method ◽  
Experimental Conditions ◽  
Reference Gene Selection ◽  
Physiological Processes ◽  
The Stability

Aspergillus is a genus of filamentous fungi with vast geographic and ecological distributions. Species within this genus are clinically, agriculturally and biotechnologically relevant, leading to increasing interest in elucidating gene expression dynamics of key metabolic and physiological processes. Reverse-transcription quantitative Polymerase Chain Reaction (RT-qPCR) is a sensitive and specific method of quantifying gene expression. A crucial step for comparing RT-qPCR results between strains and experimental conditions is normalisation to experimentally validated reference gene(s). In this review, we provide a critical analysis of current reference gene selection and validation practices for RT-qPCR gene expression analyses of Aspergillus. Of 90 primary research articles obtained through our PubMed query, 17 experimentally validated the reference gene(s) used. Twenty reference genes were used across the 90 studies, with beta-tubulin being the most used reference gene, followed by actin, 18S rRNA and glyceraldehyde 3-phosphate dehydrogenase. Sixteen of the 90 studies used multiple reference genes for normalisation. Failing to experimentally validate the stability of reference genes can lead to conflicting results, as was the case for four studies. Overall, our review highlights the need to experimentally validate reference genes in RT-qPCR studies of Aspergillus.

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