鲤鱼HSP70基因组织表达差异研究

1材料与方法 1．1材料 供试鲤鱼购自成都市洗面桥市场。主要试剂：TR—Iaol总RNA提取试剂盒购自Invitrogen公司，反转录试剂盒、pMD-18T载体、D12000DNA和如DNA聚合酶购自大连宝生物工程有限公司；琼脂糖凝胶DNA回收试剂盒购自Axygen公司；DEPC原液（Sigma分装）、JM-109感受态细胞购自大连宝生物工程有限公司，其他试剂均为国产分析纯。

Study on HSP70 Gene Expression in Different Tissue of Cyprinus carpio

Objective] The aim of this study was to investigate whether HSP70 can be used as a stress monitoring indicator in Cyprinus carpio breeding. Method] Based on HSP70 sequence of Cyprinus carpio (AY120894), one pair of primers was designed and synthesized, while the total RNA of liver tissues in Cyprinus carpio was extracted. Some cDNA fragments of Cyprinus carpio HSP70 were cloned by RT-PCR, and its differential expression in various tissues such as heart, intestine, mucus, gonad, swim bladder, gill and fin in Cyprinus carpio was also studied. Result] The cDNA sequence of 480 bp was obtained from Cyprinus carpio HSP70 gene by RT-PCR amplification. Homology comparison between the deduced amino acid sequence after sequencing and that of other types of fish showed that the homology among Cyprinus carpio, Danio rerio, Ohcorhynehus mylciss, Paralichthys olivaceus, Xiphophoorus maculates and Carassius auratus was 96%, 98%, 98%, 96%, 96% and 96% respectively. The expression of HSP70 was detected in eight tissues of Cyprinus carpio. The expression was the highest in heart, followed by swim bladder and fin, but there was no significant difference between them (P＞0.05). There was no significant difference among the expression in three tissues of intestine, mucus and fat (P＞0.05), but their expression was significantly higher than those in gonad and gill (P＜0.05). Conclusion] HSP70 gene expression is a suitable criterion for monitoring the stress degree, stress capacity and healthy conditions in Cyprinus carpio breeding.