用扣除杂交法分离藏红花苷合成相关基因的克隆

对藏红花苷合成的相关基因进行了初步分离与克隆实验研究,旨在分离与藏红花苷合成相关的特异性表达的cDNA克隆,为进一步从分子水平上研究藏红花素的合成机理奠定基础。实验采用经添加诱导子和合成前体、处于藏红花苷合成状态下的藏红花细胞和未添加诱导子和合成前体、处于生长状态的对照藏红花细胞为材料,采用扣除杂交法成功地分离到6条与藏红花苷合成相关的特异性表达的cDNA克隆。结果表明:其中一条与胡杨中由高盐环境引起的差异表达的一条mRNA的序列有较高的同源性。

Separation of gene clones related to crocin synthesis using subtractive hybridization

The crocin synthesis mechanism was investigated at the molecular level by studying the genes related to crocin syntheses. Genes controlling the translation of some key enzymes related to crocin synthesis are expressed selectively during the synthesis of crocin. Saffron (crocus sativus L.) cells begin to synthesize crocin when combined with elicitors and carotenoid precursors. Subtractive hybridization was used to compare the gene expression of naturally growing saffron cells during crocin synthesis. Six cDNA clones differentially expressed during crocin synthesis were isolated. Sequence analysis and database searches reveal that one of the six cDNA clones shows high homology to the populus euphratica mRNA differentially expressed during salt stress.