【摘要】 通过分析拟南芥与大豆的羧基转移酶α亚基的cDNA序列,运用RT PCR、RACE和基因组步行(Genomewalking)三种技术进行组合克隆,首次从油菜开花后 2 0～ 2 9天的幼胚中,克隆到质体定位的羧基转移酶α亚基的全长cDNA.同源性分析表明,其开放阅读框(ORF)编码的氨基酸序列与拟南芥、大豆的羧基转移酶α亚基的同源性分别为 85 %、5 9%.将此cDNA去除叶绿体转运肽编码序列的成熟肽编码序列,克隆到表达载体pHBM6 2 5上,蛋白质印迹分析,它在大肠杆菌中成功表达更多还原

【Abstract】 By analyzing the cDNA sequences of carboxyltransferase alpha subunits(α-CTs) from Arabidopsis thaliana and Glycine max,and combination RT-PCR, 3′-rapid amplification of cDNA ends (3′-RACE) and Genome walking techniques, the full length cDNA of plastid α-CT from embryos of Brassica napus between 20 and 29 days after flowering was cloned. Homologous analysis showed that the amino acid sequence deduced from it shares 85% and 59% homology with those of α-CT in Arabidopsis thaliana and Glycine max, respectively. The sequence encoding mature peptide that was absent from chloroplast transit peptide has been inserted into expression vector pHBM625 and expressed successfully in Escherichia coli by Western blotting analysis.更多还原