鲤鱼HSP70基因组织表达差异研究

Study on the Differential Expression of HSP70 Gene in Cyprinus carpio

[目的]研究HSP70是否可作为鲤鱼养殖中的应激监测指标。[方法]根据鲤鱼HSP70序列(AY120894)设计并合成1对引物,提取鲤鱼肝脏组织总RNA,通过RT-PCR方法克隆得到鲤鱼HSP70部分cDNA片段,然后检测其在鲤鱼心脏、肠、粘液、性腺、鳔、鳃、鳍等不同组织中的表达差异。[结果]RT-PCR扩增获得鲤鱼HSP70基因cDNA部分序列为480 bp。将测序后推测的氨基酸序列与其他种类的鱼进行同源性比较,结果发现与鲤鱼、斑马鱼、虹鳟、牙鲆、剑尾鱼、鲋鱼的同源性依次为96%、98%、98%、96%、96%、96%。HSP70在鲤鱼8个组织中均检测到表达,其中在心脏中表达最高,其次是鳔、鳍,二者之间无显著差异(P>0.05),在肠、粘液与脂肪3个组织中的表达无显著差异(P>0.05),但显著高于在性腺与鳃中的表达水平(P<0.05)。[结论]HSP70作为鲤鱼养殖中应激程度、应激能力、健康状况检测的分子标记物是可行的。

[Objective] The study was to investigate whether the heat shock protein 70（HSP70） encoding gene can be used as an indicator to monitor the stress response in carp（Cyprinus carpio） breeding.[Method] Referred to the carp HSP70 sequence（AY120894） accessed in GenBank,a pair of primers was designed to clone the cDNA fragment of HSP70 gene from the total RNA of carp liver,then the differential expressions of HSP70 gene in carp and other tissues including heart,intestine,mucus,gonad,swim bladder,gill,fin and fat,were studied.[Result] A fragment of 480 bp in length was amplified from the total RNA of carp liver via RT-PCR,the homologues of the putative amino acid sequence of this fragment and other fish species of Cyprinus carpio,Danio rerio,Ohcorhynehus mylciss,Paralichthys olivaceus,Xiphophoorus maculatus,Carassius auratus accessed in GenBank were respectively 96%,98%,98%,96%,96% and 96%.HSP70 gene was found expressed in all the eight tissues detected,of which the highest expression was observed in heart,followed by swim bladder and fin（the two are not significantly different,P〉0.05）;HSP70 gene expressed at insignificantly different level among intestine,mucus and fat（P〉0.05）,but significantly higher than that in gonad and gill（P〈0.05）.[Conclusion] It is feasible to use HSP70 gene as molecular marker to monitor the stress responsible degree,stress responsible capacity and healthy conditions of carp in breeding process.