夏威夷椰子超氧化物歧化酶基因片段的克隆与序列分析

Cloning and Sequencing of Cu·Zn-superoxide Dismutases Gene Fragment from Chamae doreacostricana L.

以热带植物夏威夷椰子(Chamaedoreacostricana)基因组DNA为模板,根据SOD基因保守序列设计特异引物进行PCR扩增,得到特异基因片段.回收该基因片段,与pMD182T载体连接,并转化到感受态大肠杆菌ER2566细胞,获得Cu·ZnSOD基因片段的克隆.序列分析表明夏威夷椰子Cu·ZnSOD基因片段含3个外显子和3个内含子,编码64个氨基酸,与玉米、红薯和白杨相应氨基酸序列的同源性分别为82.81%,81.25%,81.25%和79.69%.

Special oligonucleotides to highly conserved regions of superoxide dismutases (SOD) gene were used toprime the synthesis and amplification of gene fragment from Chamae doreacostricana L.genomic DNA by polymerase chain reaction(PCR).The fragment was linked with pMD182T vector and transformed into the competence cell of Escherichia coli ER2566 strain.After screening of recombinants,detection and sequence analysis,the transformants containing SOD gene were obtained.Sequencing analysis showed that extracted from Chamae doreacostricana Cu·Zn-SOD gene fragment,which contains three exons and three introns,had 64 nucleotides.The homologus of the amino acid sequences to those from Cryza sativa,Zea mays,Ipomoea batatas and Populus tremuloid were 82.81%,81.25%,81.25% and 79.69% respectively.