根据斑马鱼HSP90(Heat shock protein)序列(NM_131328)设计并合成一对引物。提取草鱼肝脏组织总RNA,RT—PCR扩增获得草鱼HSP90基因cDNA部分序列596bp,获得GenBank登陆号为FJ517554。将测序结果利用DNAman软件与其他几种已知序列的鱼进行比对,结果表明,草鱼HSP90与斑马鱼、鲤、鲋的同源性依次为90%、89%、92%。同时利用半定量(Semi-quantitative,SQ)RT-PCR技术检测HSP90在草鱼脂肪、肌肉、肠、脑、粘液、性腺、鳔、肝脏、心脏、脾脏、鳃、鳍12个组织的表达差异。结果表明,HSP90在草鱼除了鳍外的其他11个组织中均检测到表达,其中在心脏中表达最高,但与鳃、性腺、脑、脾脏中的表达无显著差异(P〉0.05)。在脂肪、粘液、鳔组织中的表达显著低于其他几种组织(P〈0.05),在肝脏、肌肉与肠中的表达无显著差异(P〉0.05)。
A pair of primers was designed according to zebra HSP90 (Heat shock protein)sequence(NM_131328)and total RNA was extracted from hepatopancrease of grass carp (Ctenopharyngodon idellus). 596 bp of grass carp HSP90 gene with GenBank number of FJ517554. was obtained by RT-PCR method It was found that the HSP90 gene of grass carp had 90%, 89% and 92% homolog with zebra(C, auratus) using DNAman software. The semi-quantitative (SQ RT-PCR) method was used to detect the mRNA differential expression of HSP 90 in the tissues. The results showed that the HSP90 was expressed in adipose tissue, muscle, hepatopancrease, spleen, gonad, gill, swim bladder, brain, mucilage, heart and intestine. The expression of the HSP90 in heart was more than others,but not significant difference with gill,gland,heart, brain and spleen(P〉0. 05). There was significant lower expression in adipose, mucilage,swim bladder than other tissues(P〈0.05). The expression of the HSP90 in hepatopancrease, muscle,and intestine was not siginificant.