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      저자 : Huanyu Guo (검색결과 2 건)
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        Identification of novel reference genes using sika deer antler transcriptome expression data and their validation for quantitative gene expression analysis

        Meichen Liu,Yu Zhao,Baojin Yao,Hui Zhang,Huanyu Guo,Dongyang Hu,Qun Wang 한국유전학회 2014 Genes & Genomics Vol.36 No.5

        The most commonly used normalization strategyfor quantitative real-time reverse transcription-polymerasechain reaction (RT-qPCR) is to select a stablereference gene. However, to date, no suitable referencegenes have been identified in sika deer antler tissues. Thus,the aim of this study was to identify the most stable gene ora set of genes to be used as reference genes for RT-qPCRanalysis in sika deer antler tissues. We first selected candidatereference genes using sika deer antler gene expressiondata from an Illumina sequencing platform (Hiseq2000); twenty-one reference genes from the antler tips ofChinese sika deer were selected to test for the normalizationof expression levels during different growth stages. These genes were tested by RT-qPCR and ranked accordingto the stability of their expression using two differentmethods (implemented in geNorm and NormFinder). Based on different algorithms and analytical procedures,our results clearly indicate RPL40 and Gpx as the moststable reference genes of our pool.

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        Preparation and molecular characterization of a polyclonal antibody as an efficient cutworm reference protein

        Huan Yu,Lei Hea,Yi-Yi Ou-Yang,Guo-Hua Huang 한국응용곤충학회 2018 Journal of Asia-Pacific Entomology Vol.21 No.3

        Antibodies, which are wildly used in molecular researches, are proteins secreted by plasma cells that can specifically recognize specific antigens. To obtain a stable reference antibodies are important for protein analysis. In this study, a capable glyceraldehyde phosphate dehydrogenase (GAPDH) antiserum against most noctuid larval protein samples was prepared. Firstly, the full-length gapdh was amplified from Spodoptera exigua larvae to construct the prokaryotic expression vector. The purified His-tag fused GAPDH protein was used to immunize rabbits for the antiserum preparation. Protein samples extracted from 11 insect species distributed across seven families and three orders were used to perform the Western bolt analysis. The bright specific bands were detected in S. exigua, S. litura, Helicoverpa armigera, and Mythimna seperata protein samples, indicating that this antiserum may be capable of detecting noctuid larval encoded GAPDH. Further immunofluorescence identification of H. armigera and S. exigua paraffin section slides with GAPDH antiserum exhibited an identical cytostained image. The GAPDH antiserum prepared in this study is useful and efficient as reference antibodies in studies involving noctuid larval protein samples in other related fields.

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