泥鳅elovl1基因克隆、表达分析及其在低温胁迫下的响应
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国家自然科学基金面上项目(31672660)


Cloning and characteristic analysis of elovl1 genes from loach (Misgurnus anguillicaudatus) and its role in adaptation to cold temperature
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    摘要:

    为探究泥鳅ELOVL1的功能和作用,采用RT-PCR和RACE方法克隆泥鳅的elovl1基因,并分析elovl1基因的表达规律,最后将泥鳅分别暴露在26℃和8℃下饲养30 d,研究低温胁迫对泥鳅脂肪酸组成、组织形态及elovl1基因表达的影响。结果发现:elovl1a全长为2 216 bp,ORF为948 bp,编码315个氨基酸;elovl1b全长为1 895 bp,ORF为963 bp,编码320个氨基酸。氨基酸序列特征如下:1个高保守的氧化还原组氨酸簇(HXXHH),多个跨膜区,氨基酸序列C端均含有内质网滞留信号(KXKXX)。elovl1a和elovl1b在所有组织中表达,elovl1a在每个组织的表达量均低于elovl1b。低温下总SFA明显降低(P<0.05),总MUFA显著增加(P<0.05)。低温显著诱导elovl1a、elovl1b表达。研究结果表明:泥鳅ELOVL1在脊椎动物中保守,ELOVL1可能在机体适应低温时,参与产生能量,起到重要的作用。

    Abstract:

    This trial was conducted to investigate the role of the elovl1 gene from loach (Misgurnus anguillicaudatus).The loach elovl1 was cloned by using RT-PCR and RACE,and its expression patterns were analyzed.The loach were exposed to 8℃ and 26℃ for 30 d to evaluate the effects of cold stress on fatty acid composition,tissue morphology and expression of the elovl1 gene. The results showed that the elovl1a full-length cDNA was 2 216 bp with an ORF of 948 bp,encoding 315 amino acids and the elovl1b full-length cDNA was 1 895 bp with an ORF of 963 bp,encoding 320 amino acids.The elovl1 protein was conserved in vertebrates,containing one conserved diagnostic histidine box HXXHH motif,several transmembrane regions,and the endoplasmic reticulum retrieval signal (KXKXX) at the carboxyl terminus.The elovl1a and elovl1b were expressed in almost all tissues and the expression of elovl1a was lower than that of elovl1b in every tissue tested.Total saturated fatty acids (SFA) significantly decreased (P<0.05),and total monounsaturated fatty acids (MUFA) significantly increased in the liver at 8℃ compared to that at 26℃ (P<0.05).Both elovl1a and elovl1b were significantly induced in the muscle and liver (P<0.05) after exposed to cold.In conclusion,elovl1 was conserved in vertebrates,which might involve in energy production and play an important role in adaptation to cold temperature.

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张义凤,王丽伟,曹小娟,高坚.泥鳅elovl1基因克隆、表达分析及其在低温胁迫下的响应[J].华中农业大学学报,2018,37(04):102-111

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  • 在线发布日期: 2018-07-09
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