【摘要】 根据已报道的植物3-羟基,3-甲基戊二单酰辅酶A还原酶(HMGR)和环阿屯醇合成酶(CS)基因保守序列分别合成简并引物,先后从盾叶薯蓣(Dioscorea zingiberensis)幼苗中克隆了一段663 bp(HMGR-DZh)和一段1 245 bp(CS-DZc)的cDNA片段,经NCBI和CLUSTALW在线序列同源分析,结果表明,由HMGR-DZhcDNA推导的蛋白质与已报道植物HMGR氨基酸序列相似性达到63%~70%.由CS-DZc cDNA推导的蛋白质氨基酸序列中具有环氧角鲨烯环化酶(OSCs)特异保守结构域:DCTAE结构域和C末端的2个QW高度保守区,并与已报道植物环阿屯合成酶氨基酸序列相似性达到80%以上.本实验还分别以HMGR-DZh和CS-DZccDNA为探针进行了Southern和Northern杂交分析.Northern杂交分析表明,盾叶薯蓣HMGR-DZh基因的mRNA约为1.8~2 kb,CS-DZc基因的mRNA约为2~2.5 kb.更多还原

【Abstract】 A 663 bp cDNA fragment(HMGR-DZh) and a 1 245 bp cDNA(CS-DZc) fragment were cloned from the seeding of Dioscorea zingiberensis by RT-PCR method.Sequencing analyses by NCBI blast and CLUSTALW alignment showed that,the HMGR-DZh protein had 63%～70% identity with HMGR protein of other plants reported.The comparability of CS-DZc protein to CS protein of other plants reported was up to 80%.The DCTAE and two QW high conservative domain,which was characteristic in CS protein,were found in the C tag of CS-DZc.Northern blotting has showed that the full-length mRNA of HMGR-DZh is about 1.8～2.0 kb and CS-DZc is about 2.0～2.5 kb.更多还原