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黃鱔PLA2基因的克隆及表達(dá)分析

發(fā)布時(shí)間:2018-02-23 21:13

  本文關(guān)鍵詞: 磷脂酶A 黃鱔(Monopterus albus) 克隆 組織表達(dá) 出處:《淡水漁業(yè)》2017年03期  論文類型:期刊論文


【摘要】:利用同源克隆技術(shù)和RACE技術(shù)克隆黃鱔(Monopterus albus)磷脂酶A2(PLA2)基因的cDNA全長;并采用熒光定量RT-PCR法檢測PLA2基因在黃鱔各組織中的表達(dá)量情況。結(jié)果顯示:黃鱔PLA2基因全長cDNA大小為2 606 bp(Gen Bank登錄號:KX852397),其中開放閱讀框2 205 bp,編碼736個(gè)氨基酸,預(yù)測分子大小為83.623 kD,理論等電點(diǎn)5.84;5'和3'非編碼區(qū)長度分別為208 bp和193 bp。該蛋白序列沒有信號肽和跨膜結(jié)構(gòu)。氨基酸序列比對和系統(tǒng)樹分析顯示,黃鱔PLA2基因的結(jié)構(gòu)十分保守,黃鱔與大黃魚和金頭鯛的PLA2基因親緣關(guān)系最近,與鼠類和蟾蜍的親緣關(guān)系較遠(yuǎn)。RT-PCR分析表明,PLA2基因在組織中的表達(dá)具有組織特異性,在消化器官前腸組織中的表達(dá)最高,在性腺及肌肉組織中表達(dá)很少。
[Abstract]:The full-length cDNA of Monopterus albusus phospholipase A2PLA2) gene was cloned by homologous cloning technique and RACE technique. The expression of PLA2 gene in different tissues of Monopterus Albus was detected by fluorescence quantitative RT-PCR. The results showed that the full-length cDNA of PLA2 gene of Monopterus Albus was 2 606 bp(Gen Bank accession number: KX852397, and the open reading frame 2 205 BP encoded 736 amino acids. The predicted molecular size is 83.623 KD, the theoretical isoelectric point 5.84 'and the length of 3' noncoding region are 208 BP and 193 BP, respectively. The protein sequence has no signal peptide and transmembrane structure. Amino acid sequence alignment and phylogenetic tree analysis show that the structure of PLA2 gene of Monopterus Albus is very conserved. The relationship between PLA2 gene of Monopterus Albus and Pseudosciaena Crocea and Sparus macrocephalus was close. RT-PCR analysis showed that the expression of PLA2 gene was tissue specific in tissues and the highest in the tissues of digestive organs. There is little expression in the gonads and muscles.
【作者單位】: 江西農(nóng)業(yè)大學(xué)動(dòng)物科學(xué)技術(shù)學(xué)院;江西省水產(chǎn)科學(xué)研究所;
【基金】:國家自然科學(xué)基金(31360641)
【分類號】:S917.4
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本文編號:1527626

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