淞江鲈HSP70基因cDNA部分序列克隆及其组织表达差异  

Cloning of Partial HSP70 cDNA Sequence and its Tissue Differential Expression in Rough-skinned Sculpin

作　　者：刘庆全[1] 罗武松[1] 郑映荷[1] 许耀[1] 高峰 秦志浩 倪雯雯[1] 王金秋[1] 

机构地区：[1]复旦大学生命科学学院,遗传学研究所,遗传学国家重点实验室,上海200433 [2]上海四鳃鲈水产科技发展有限公司,上海200433

出　　处：《复旦学报（自然科学版）》2013年第4期558-563,共6页Journal of Fudan University：Natural Science

基　　金：上海市农委科技兴农重点攻关项目(沪农科攻字(2010)第1-7-2号);农业部公益性行业(农业)科研专项(201203065)

摘　　要：为了研究HSP70(heat shock protein 70)是否可作为淞江鲈(Trachidermus fasciatus Heckel)养殖中的应激监测分子指标,根据鲤鱼热应激蛋白70基因序列(AYl20894)设计并合成引物,以淞江鲈肝脏组织总RNA为模板,获得淞江鲈HSP70基因cDNA部分序列,长度为480bp,GenBank登陆号为KC342053.序列分析结果显示此序列与红笛鲷(Lutjanus sanguineus)等有较高的同源性.同时,应用所获淞江鲈HSP70基因部分序列,以淞江鲈β-actin基因的序列(HM449124)为内参,利用荧光定量PCR技术,检测淞江鲈肌肉、肠、脑、皮肤、性腺、肝脏、心脏、鳃、鳍9个组织的表达差异.结果表明,HSP70在淞江鲈9个组织中普遍表达,其中在鳍中表达量最高,在性腺、鳃、脑、肌肉、肝脏、皮肤、肠中的表达量依次递减,在心脏中表达量最低.在鳍中HSP70的表达量可能成为衡量淞江鲈应激程度的指标.To determine whether HSP70 could be utilized as an indicator for stress detection in Rough-skinned Sculpin（Trachidermusfasciatus Heckel） cultivation, the gene＇s partial sequence of 480 bp（GenBank accession number： KC342053） was amplified with total RNA extracted from adult Rough-skinned Sculpin as model and primers designed based on the sequences of carp HSP70 sequence（AY120894）. Sequence analysis indicated this sequence shares high similarity with its counterpart in Lutjanus sanguineus. Meanwhile, Rough-skinned Sculpin HSP70 fluorescence quantitative PCR platform was established with primers based on the partial sequence obtained and the Rough-skinned Sculpin fl-actin sequence（HM449124）. The expression patterns of HSPT0 in nine different tissues namely muscle, intestine, brain, skin, gonad, liver, heart, gill and fin were detected. The results demonstrated that HSP70 expressed in all the nine tissues, among which the expression in the fin was of the most intensity. The seven tissues of gonad, gill, brain, muscle, liver, skin, intestine were in a descending order concerning the expression intensity of HSP70, while the heart had the lowest level of HSP70 expression, suggesting the expression level of HSP70 in fin could be used as an indicator.

关 键 词：热应激蛋白70 克隆 组织差异 荧光定量 淞江鲈 

分 类 号：S917[农业科学—水产科学] Q786[生物学—分子生物学]