甘蓝型油菜BnCHI-1基因的克隆和表达特征分析

发布时间：2018-01-01 15:13

  本文关键词：甘蓝型油菜BnCHI-1基因的克隆和表达特征分析　出处：《中国油料作物学报》2016年02期 　论文类型：期刊论文

  更多相关文章： 甘蓝型油菜 BnCHI-基因 克隆 表达 查尔酮异构酶

【摘要】：为了解Bn CHI-1基因的表达与粒色差异之间的关系,采用RACE技术从甘蓝型油菜黑籽系5B中克隆了Bn CHI-1全长c DNA序列和基因组序列。Bn CHI-1的DNA序列为1 809bp。965bp的Bn CHI-1 mRNA含有一个756bp的ORF,编码包含251个氨基酸的多肽。预测发现Bn CHI-1蛋白存在查尔酮-黄烷酮异构酶结构域,底物2S-柚皮苷的结合位点以及氢键网络氨基酸残基活性位点。Southern杂交结果表明在甘蓝型油菜中有6个CHI基因成员。RT-PCR揭示,甘蓝型油菜5B的11个器官组织均有Bn CHI转录,其在茎、叶、蕾和花中的转录水平比较高,其次为种子和根,在子叶中最低。比较1对近等基因系(黑籽系L1和黄籽系L2)中Bn CHI转录水平,发现在花蕾、花、花后20d种子和花后30d种子中,二者的转录水平没有显著差异,但在开花后10d的种子中,L2中Bn CHI转录水平显著低于L1中Bn CHI转录水平。
[Abstract]:In order to understand the relationship between the expression of Bn CHI-1 gene and the difference of granulocyte color. The full length c DNA sequence of Bn CHI-1 and the DNA sequence of genome. Bn CHI-1 were cloned from the black seed line 5B of Brassica napus by RACE technique. The BP CHI-1 mRNA of 809bp.965bp contained a 756bp ORF. Bn CHI-1 protein was predicted to have a chalcone-flavanone isomerase domain. The binding site of substrate-2S-naringin and the activity site of amino acid residues of hydrogen bond network. Southern blot analysis showed that there were six CHI gene members. RT-PCR in Brassica napus. Bn CHI transcription was found in 11 organs of Brassica napus 5B. The transcription level was higher in stems, leaves, buds and flowers, followed by seeds and roots. The transcriptional levels of Bn CHI in 1 pair of near-isogenic lines (black seed line L1 and yellow seed line L2) were compared. The results showed that the transcription level of Bn CHI was found in bud, flower, seed 20 days after anthesis and seed 30 days after anthesis. There was no significant difference in transcription level between the two groups, but the Bn CHI transcription level in L2 was significantly lower than that in L1 at 10 days after anthesis.
【作者单位】： 长江大学生命科学学院;西南大学农学与生物科技学院;中国农业科学院油料作物研究所;
【基金】：国家自然科学基金(31171177)
【分类号】：S565.4
【正文快照】： 查尔酮异构酶(chalcone isomerase,CHI,EC5.5.1.6)是类黄酮代谢途径中的一个关键酶,它催化4,2’,4’,6’-四羟基查尔酮(naringenin chal-cone)形成黄烷酮。虽然该反应在自然界中可以自发发生,但CHI可以使反应速度提高107倍[1],CHI也对(2S)-黄烷酮的生化活性有重要影响[2,3]。，

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