黄鳝PLA2基因的克隆及表达分析

发布时间：2018-02-23 21:13

  本文关键词： 磷脂酶A 黄鳝(Monopterus albus) 克隆 组织表达　出处：《淡水渔业》2017年03期 　论文类型：期刊论文

【摘要】：利用同源克隆技术和RACE技术克隆黄鳝(Monopterus albus)磷脂酶A2(PLA2)基因的cDNA全长;并采用荧光定量RT-PCR法检测PLA2基因在黄鳝各组织中的表达量情况。结果显示:黄鳝PLA2基因全长cDNA大小为2 606 bp(Gen Bank登录号:KX852397),其中开放阅读框2 205 bp,编码736个氨基酸,预测分子大小为83.623 kD,理论等电点5.84;5'和3'非编码区长度分别为208 bp和193 bp。该蛋白序列没有信号肽和跨膜结构。氨基酸序列比对和系统树分析显示,黄鳝PLA2基因的结构十分保守,黄鳝与大黄鱼和金头鲷的PLA2基因亲缘关系最近,与鼠类和蟾蜍的亲缘关系较远。RT-PCR分析表明,PLA2基因在组织中的表达具有组织特异性,在消化器官前肠组织中的表达最高,在性腺及肌肉组织中表达很少。
[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.
【作者单位】： 江西农业大学动物科学技术学院;江西省水产科学研究所;
【基金】：国家自然科学基金(31360641)
【分类号】：S917.4
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本文编号：1527626