人铜锌超氧化物歧化酶cDNA的克隆与测序

Cloning and sequencing of Cu,Zn superoxide dismutase cDNA in human

目的探讨通过分子生物学技术克隆人铜锌超氧化物歧化酶的cDNA片段。方法从人正常肝细胞中提取总RNA,然后对其进行反转录产生第1条cDNA链。根据GeneBank中人铜锌超氧化物歧化酶(Cu/Zn-SOD)cDNA序列(序列号AB087266)设计1对PCR引物,对反转录后产生的第1条cDNA链进行PCR扩增,获得目的基因。结果获得459bp预期大小的PCR产物,经琼脂糖凝胶电泳和测序验证,使用序列分析软件证明,提取的是所需要的hSOD1cDNA的片段。结论获得的目的片段纯度高,未降解,能满足进一步试验的需要。

Objective To clone of Cu/Zn superoxide dismutase cDNA fragment in human by molecular biology techniques.Methods The total RNA was extracted from human liver,and then reversed transcriptase it first chain.According to Gene Bank,human Cu/Zn-SOD cDNA sequence（serial number AB087266）,we designed a pair of PCR primers,and generated the first PCR cDNA chain after reverse transcription and then obtained gene.Results Agarose gel electrophoresis,sequenced by sequence analysis software showed that the expected size of 459bp PCR product was the hSOD1cDNA fragment which was required.Conclusion The purpose gene fragments is high purity,without degradation,and it is used to further study.