本文關(guān)鍵詞：甘藍型油菜BnCHI-1基因的克隆和表達特征分析　出處：《中國油料作物學(xué)報》2016年02期 　論文類型：期刊論文

  更多相關(guān)文章： 甘藍型油菜 BnCHI-基因 克隆 表達 查爾酮異構(gòu)酶

【摘要】：為了解Bn CHI-1基因的表達與粒色差異之間的關(guān)系,采用RACE技術(shù)從甘藍型油菜黑籽系5B中克隆了Bn CHI-1全長c DNA序列和基因組序列。Bn CHI-1的DNA序列為1 809bp。965bp的Bn CHI-1 mRNA含有一個756bp的ORF,編碼包含251個氨基酸的多肽。預(yù)測發(fā)現(xiàn)Bn CHI-1蛋白存在查爾酮-黃烷酮異構(gòu)酶結(jié)構(gòu)域,底物2S-柚皮苷的結(jié)合位點以及氫鍵網(wǎng)絡(luò)氨基酸殘基活性位點。Southern雜交結(jié)果表明在甘藍型油菜中有6個CHI基因成員。RT-PCR揭示,甘藍型油菜5B的11個器官組織均有Bn CHI轉(zhuǎn)錄,其在莖、葉、蕾和花中的轉(zhuǎn)錄水平比較高,其次為種子和根,在子葉中最低。比較1對近等基因系(黑籽系L1和黃籽系L2)中Bn CHI轉(zhuǎn)錄水平,發(fā)現(xiàn)在花蕾、花、花后20d種子和花后30d種子中,二者的轉(zhuǎn)錄水平?jīng)]有顯著差異,但在開花后10d的種子中,L2中Bn CHI轉(zhuǎn)錄水平顯著低于L1中Bn CHI轉(zhuǎn)錄水平。
[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.
【作者單位】： 長江大學(xué)生命科學(xué)學(xué)院;西南大學(xué)農(nóng)學(xué)與生物科技學(xué)院;中國農(nóng)業(yè)科學(xué)院油料作物研究所;
【基金】：國家自然科學(xué)基金(31171177)
【分類號】：S565.4
【正文快照】： 查爾酮異構(gòu)酶(chalcone isomerase,CHI,EC5.5.1.6)是類黃酮代謝途徑中的一個關(guān)鍵酶,它催化4,2’,4’,6’-四羥基查爾酮(naringenin chal-cone)形成黃烷酮。雖然該反應(yīng)在自然界中可以自發(fā)發(fā)生,但CHI可以使反應(yīng)速度提高107倍[1],CHI也對(2S)-黃烷酮的生化活性有重要影響[2,3]。，

本文編號：1365031