论文部分内容阅读
首先以一个用cDNA-AFLP分析大白菜[Brassica campestris L.ssp.pekinensis(Lour.)Olsson]雄性不育花蕾败育得到的差异表达片段(TDF,Transcript Derived Fragments)EST54-1为信息探针,在Gen-Bank数据库中进行同源EST序列检索,并对亲缘关系近的同源EST序列进行拼接,获得长度为1574bp的虚拟序列。然后设计引物,经RT-PCR扩增、测序,获得了大白菜苹果酸脱氢酶基因的cDNA全长序列,并已将其登录到GenBank,登录号:FJ208590,该cDNA全长1209bp,编码403个氨基酸。同源分析显示该cDNA与电子克隆获得的核酸序列99%同源,该cDNA序列推导的氨基酸序列与拟南芥苹果酸脱氢酶同源性100%。虽然在大白菜花蕾败育过程中苹果酸脱氢酶基因mRNA表达量下降,但其与大白菜花蕾败育的关系还需通过RNA干扰等技术进行验证。
First, a cDNA-AFLP analysis of Transgenic Derived Fragments (TDF) EST54-1 was used as informative probe to analyze the abortion of Chinese cabbage [Brassica campestris L.ssp.pekinensis (Lour.) Olsson] The homologous EST sequence was searched in Gen-Bank database, and the homologous EST sequences close relatives were spliced to obtain a virtual sequence with a length of 1574bp. Then, the full length cDNA of malate dehydrogenase gene was obtained by RT-PCR amplification and sequencing. The full-length cDNA of malate dehydrogenase gene was obtained and registered in GenBank with accession number FJ208590. The full-length cDNA was 1209bp encoding 403 Amino acids. Homology analysis showed that the cDNA was 99% homologous to that obtained from the electronic cloning. The amino acid sequence deduced from the cDNA sequence was 100% homologous to Arabidopsis malate dehydrogenase. Although the mRNA expression of malate dehydrogenase gene was decreased in the process of flower bud abortion in Chinese cabbage, its relationship with flower bud abortion in Chinese cabbage needs to be verified by RNA interference and other techniques.