| 王修芳，劉慶慧，吳垠，黃倢.中國明對蝦（Fenneropenaeus chinensis）coat-ε基因全長cDNA尅隆及組織分佈.漁業科學進展，2016,37（4）：147-152 |
| 中國明對蝦（Fenneropenaeus chinensis）coat-ε基因全長cDNA尅隆及組織分佈 |
| cDNA Cloning of Coat-Epsilon Gene and Its Tissue Distribution in Fenneropenaeus chinensis |
| 投稿時間：2015-06-17 修訂日期：2015-07-29 |
| DOI：10.11758/yykxjz.20150617002 |
| 中文關鍵字： 中國明對蝦 coat-ε 基因克隆 組織分佈 |
| 英文關鍵字： Fenneropenaeus chinensis Coat-ε Gene clone Tissue distribution |
| 基金項目：國家重點基礎研究發展計畫（2012CB114401）、泰山學者“建設工程專項經費”和農業部科研傑出人才和創新團隊專項經費共同資助 |
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| 摘要點擊次數： 4612 |
| 全文下載次數： 4508 |
| 中文摘要： |
| coat-ε基因表達的蛋白是組成COPⅠ的coatomer複合體的一個亞基，為獲得中國明對蝦（Fenneropenaeus chinensis）coat-ε基因全長序列，採用cDNA末端快速擴增（Rapid amplification of cDNA end，RACE）科技，擴增出coat-ε基因的3¢端和5¢端，測序結果經DNAMAN比對拼接得出coat-ε基因全長，基因全長1402 bp，5¢非編碼區（UTR）84 bp，3¢非編碼區（UTR）310 bp，開放閱讀框1008 bp，預測編碼335個胺基酸，其中，第230–300的胺基酸屬於TPR超家族，SignalP 3.0 Server預測胺基酸序列沒有信號肽，TMHMM Server v. 2.0分析此胺基酸不存在跨膜結構，PSORTⅡPrediction預測該蛋白位於線粒體、細胞質、內質網中，屬胞內蛋白。系統進化樹顯示，中國明對蝦的coat-ε基因與節肢動物門的動物親緣關係相近。採用實时螢光定量方法分析該基因在鰓、上皮、胃、肌肉、肝胰腺等不同組織中的相對表達，結果顯示，coat-ε在肌肉中的相對轉錄表達量最高，在鰓和附肢的表達次之。本研究獲得的中國明對蝦coat-ε全長序列，可為該基因功能研究提供基礎。 |
| 英文摘要： |
| Coat-epsilon protein（coat-ε）is a subunit of the coatomer complex that forms COPⅠ.To obtain the full-length sequence of coat-εof Fenneropenaeus chinensis，we first acquired the sequences of 3' and 5' ends using rapid amplification of cDNA ends（RACE）.The results were then spliced by DNAMAN to obtain the full-length 1402 bp sequence.The predicted 5' non-coding region（UTR）had 84 bp and the 3' non-coding region（UTR）had 310 bp.The open reading frame had 1008 bp that was supposed to encode 335 amino acids.The fragment including 230aa to 300aa belonged to the TPR superfamily.Signalp 3.0 server and TMHMM Server Version 2.0 analysis suggested that the amino acid sequence did not contain a signal peptide or a transmembrane structure.PSORTⅡPrediction showed that coat-epsilon was probably located in mitochondria，cytoplasm，and endoplasmic reticulum.The phylogenetic tree analysis showed that coat-εof F. chinensis was closely related to that of Arthropoda.We also analyzed the mRNA expression of coat-εin different tissues with quantitative real-time PCR，and found that it was expressed in all tested tissues including appendage，hepatopancreas，epithelium，heart，stomach，intestine，eyestalk，muscles，gill，lymphoid organ and hemocytes.The expression level was the highest in muscles，followed by the gill and appendage.Our results provided important information for the functional study of coat-ε. |
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