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Position: Home > Articles > Cloning and Sequence Analyzing of Malate Dehydrogenase Gene in Chinese Cabbage [Brassica campestris L.ssp.pekinensis (Lour.) Olsson] Acta Horticulturae Sinica 2009,36 (3) 363-368

大白菜苹果酸脱氢酶基因的克隆及序列分析

作  者：
贾晋;张鲁刚
单  位：
西北农林科技大学园艺学院;内蒙古科技大学生物与化学工程学院
关键词：
大白菜;花蕾败育;电子克隆;RT-PCR;苹果酸脱氢酶
摘  要：
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。
译  名：
Cloning and Sequence Analyzing of Malate Dehydrogenase Gene in Chinese Cabbage [Brassica campestris L.ssp.pekinensis (Lour.) Olsson]
作  者：
JIA Jin1,2 and ZHANG Lu-gang1(1 College of Horticulture,Northwest A & F University,Yangling,Shaanxi 712100,China;2School of Biological & Chemical Engineering,Inner Mongolia University of Science & Technology,Baotou,Inner Mongolia 014010,China)
关键词：
Chinese cabbage;bud aborting;in silico cloning;RT-PCR;malate dehydrogenase
摘  要：
Firstly,a differential TDF(Transcript Derived Fragments)EST54-1 from the RNA of bud aborting of male sterile Chinese cabbage by cDNA-AFLP analysis was used as a querying probe to blast homologous ESTs sequences in the GenBank database,we spliced the homologous EST sequences assembled from GenBank,and gained an virtual sequence of 1 574 bp length.Then,two primers were designed from it,the full length cDNA of malate dehydrogenase gene in Chinese cabbage was gained through RT-PCR amplifying and sequencing and entered GenBank as accession number FJ208590,which is 1 209 bp length and coded 403 amino acids.Homologous analysis showed that the cDNA amplified had 99% identity with sequence from in silico cloning on the nucleotide acid sequence,and amino acids sequence derived from it had 100% identity with Malate Dehydrogenase(MDH)in Arabidopsis.Though the expression capacity of MDH gene in Chinese cabbage was depressed as bud aborting,the relationship between MDH and bud abortion in Chinese cabbage need to be clarified through further research such as RNAi technology etc.

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