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金乌贼热休克蛋白基因的挖掘及对低盐胁迫的响应
王兴强1, 郑年昊1, 崔春辉1, 曹 梅1, 沈 晔1, 刘 莹1, 张子文1, 汤子威1, 秦传新2, 陈百尧3
1.江苏海洋大学海洋生命与水产学院;2.中国水产科学研究院南海水产研究所;3.连云港市农业农村局
摘要:
热休克蛋白(HSPs)为动植物响应外界胁迫产生的一类蛋白质,能有效改善机体对外界胁迫的适应能力。本研究基于先前获得的低盐胁迫金乌贼高通量转录组数据,对与低盐胁迫密切相关的HSPs(HSP和crystallin家族)基因进行挖掘;鉴于实时定量内参基因筛选,同时挖掘出金乌贼肌动蛋白(actin)家族系列基因;然后,对actin、HSP和crystallin家族基因及其编码的氨基酸序列进行生物信息学分析并探究低盐胁迫对金乌贼HSP和crystallin家族基因表达的影响。金乌贼actin家族基因包括actin、内收肌actin、胞质actin、actin 1、actin II和β actin 6个成员,HSP家族基因包括HSP β、小HSP、HSP 10、HSP 16、HSP 30、HSP 60、HSP 70、HSP 75、HSP 90α、HSP 90和HSP 90β 11个成员,金乌贼crystallin家族基因包括S crystallin、S crystallin 6、S crystallin SL18和O crystallin 4个成员。氨基酸序列比对发现,actin家族6个基因共享甘氨酸、精氨酸、丝氨酸、异亮氨酸、苏氨酸和天冬氨酸等13个位点,crystallin家族4个基因共享苯丙氨酸、天冬酰胺、甘氨酸、天冬氨酸、脯氨酸、精氨酸、酪氨酸、丝氨酸和半胱氨酸等17个位点,而HSP家族HSP β、小HSP、HSP 10、HSP 16和HSP 30 5个基因共享1个甘氨酸位点和4个保守位点,HSP 60、HSP 70、HSP 75、HSP 90α、HSP 90和HSP 90β 6个基因仅共享2个保守位点。实时定量结果发现,低盐胁迫可以明显提高金乌贼HSP β、HSP 16、HSP 30和HSP 70的基因表达,而明显降低crystallin家族基因的表达,表明在低盐胁迫条件下,金乌贼HSPs基因通过自身的差异表达来提高机体对外界胁迫的适应防御能力。
关键词:  金乌贼  低盐胁迫  肌动蛋白  热休克蛋白  晶体蛋白
DOI:
分类号:Q768
基金项目:广东省渔业生态环境重点实验室开放基金(FEEL_2019_3); 连云港市海燕计划科研项目
Excavation of heat shock protein gene and its response to low salinity stress in Sepia esculenta
WANG Xing-qiang1, ZHENG Nian-hao1, CUI Chun-hui1, CAO Mei1, SHEN Ye1, LIU Ying1, ZHANG Zi-wen1, TANG Zi-wei1, QIN Chuan-xin2, CHEN Bai-yao3
1.College of Marine Life and Fisheries,Jiangsu Ocean University;2.South China Sea Fisheries Research Institute,Chinese Academy of Fisheries Sciences;3.Lianyungang Agricultural and Rural Bureau
Abstract:
Heat shock proteins (HSPs) are a kind of proteins produced by plants and animals in response to external stress and can effectively improve the adaptability of the body to external stress. In this study, based on the previously obtained high-throughput transcriptome data of Sepia esculenta under low salinity stress, HSPs (HSP and crystallin family) genes closely related to low salinity stress were mined. In view of real-time quantitative internal reference gene screening, actin family genes of S. esculenta were mined. Then, the actin, HSP and crystallin family genes and their encoded amino acid sequences were analyzed by bioinformatics and the effect of low salinity stress on the gene expression of HSP and crystallin family of S.esculenta was explored. The actin family genes of S.esculenta include actin, adductor actin, cytoplasmic actin, actin 1, actin II and β actin. HSP family genes include 11 members of HSP β, small HSP, HSP 10, HSP 16, HSP 30, HSP 60, HSP 70, HSP 75, HSP 90 α, HSP 90 and HSP 90 β. The crystallin family gene of S.esculenta includes four members: S crystallin, S crystallin 6, S crystallin SL18 and O crystallin. Amino acid sequence alignment showed that six genes of actin family shared 13 loci of glycine, arginine, serine, isoleucine, threonine and aspartic acid, while four genes of crystallin family shared 17 loci of phenylalanine, asparagine, glycine, aspartic acid, proline, arginine, tyrosine, serine and cysteine. HSP β, small HSP, HSP 10, HSP 16 and HSP 30 shared one glycine locus and four conserved loci, while HSP 60, HSP 70, HSP 75, HSP 90 α, HSP 90 and HSP 90 β only shared two conserved loci. The real-time quantitative results showed that under low salinity stress, HSP β, HSP16, HSP30 and HSP70 gene expression of S.esculenta were significantly increased, while the expression of crystallin family gene was significantly decreased, which indicated that HSPs gene of S.esculenta could improve the adaptive defense ability to external stress through its own differential expression.
Key words:  Sepia esculenta  low salinity stress  actin  heat shock protein  crystalline
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