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本研究使用生物信息学相关软件对本课题组前期获得的枣树2-半胱氨酸氧化还原酶基因Zj2-CP(Gen Bank登录号:AB812086)的c DNA序列进行了分析。结果表明:该序列是一个全长为834 bp的开放阅读框,可编码277个氨基酸,分子质量为30.65 k D,理论等电点8.95,无信号肽和跨膜区,不属于分泌蛋白,有亲水性;二级结构以无规则卷曲为主,三级结构成弯曲螺旋状。氨基酸序列相似性分析表明,Zj2-CP基因编码的氨基酸序列与豇豆、玉米氨基酸序列的同源性最高,达到90%。以Zj2-CP基因的c DNA序列为模板进行PCR扩增,用内切酶消化后与载体PEZR(K)-LNY连接并转入大肠杆菌感受态细胞DH5α中,经酶切及测序证明,植物表达载体构建成功。研究结果可为深入探讨枣树2-半胱氨酸氧化还原酶基因的功能及枣树抗逆机制提供基础。
In this study, c DNA sequences of Zj2-CP (GenBank accession number: AB812086) of jujube tree 2-cysteine oxidoreductase gene obtained in our group were analyzed by bioinformatics software. The results showed that the sequence was an open reading frame (ORF) of 834 bp in length, encoding 277 amino acids with a molecular mass of 30.65 kD and a theoretical isoelectric point of 8.95. The sequence contained no signal peptide and transmembrane domain and did not belong to secreted protein. Hydrophilic; Secondary structure with random curl-based, three-stage structure into a curved spiral. Amino acid sequence similarity analysis showed that the amino acid sequence encoded by Zj2-CP gene had the highest homology with cowpea and corn amino acid sequences, reaching 90%. The cDNA of Zj2-CP gene was amplified by PCR and ligated with vector PEZR (K) -LNY after digested with endonuclease and transferred into DH5α of E.coli competent cells. After digestion and sequencing, The expression vector was constructed successfully. The results can provide a foundation for further exploring the function of 2-cysteine oxidoreductase gene and jujube stress-resistance mechanism.