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象草CCR基因的克隆与生物信息学分析(英文)
  • ISSN号:1009-4229
  • 期刊名称:Agricultural Science&technology
  • 时间:0
  • 页码:284-291
  • 分类:S[农业科学]
  • 作者机构:[1]华南农业大学农学院,广东广州510642
  • 相关基金:国家自然科学基金项目(30972138);广东省自然科学基金项目(9451064201003804).
  • 相关项目:象草4CL基因的生物信息学分析与表达调控研究
中文摘要:

[目的]从能源植物象草中克隆出参与木质素生物合成的肉桂酰辅酶A还原酶基因(CCR)的cDNA序列及全长DNA序列,进而分析其序列特征。[方法]采用传统RT-PCR及RACE技术克隆象草CCR序列,并利用NCBI,ProtParam,ProtScale,TMHMM,TargetP,SignalP,Pfam20.0,Prosite,Swiss-Model和ClustalW2等在线分析程序以及DNAman,DNAstar和MEGA5软件对得到的序列进行生物信息学分析。[结果]克隆得到了象草CCR包含编码区和3'非翻译区的长为1316bp的cDNA序列以及包含5个外显子和4个内含子的全长为6133bp的DNA序列。通过生物信息学分析得知,象草CCR基因编码长为369个氨基酸的蛋白,该蛋白二级结构元件以无规则卷曲和α-螺旋为主,属于依赖NAD表异构酶/脱氢酶家族,其辅助因子结合区域和底物结合位点高度保守。[结论]成功从象草中克隆出肉桂酰辅酶A还原酶基因。它具有CCR同源基因典型的特征。获得的各种生物信息学数据为今后开展此酶的深入研究以及对象草的更好利用提供了一定的理论参考价值。

英文摘要:

[Objective] The aim was to clone the cDNA and DNA sequences of the CCR (Cinnamoyl-CoA reductase) gene which involves in lignin biosynthesis, from Pennisetum purpureum, and to make comprehensive analysis on these sequences. [Method] CCR sequences were cloned from P. purpureum by using conventional RT-PCR and RACE (Rapid Amplification of cDNA Ends) methods; and the bioinformatic analyses of the CCR were conducted by means of NCBI, ProtParam ProtScale, TMHMM, TargetP, SignalP, Pfam20.0, Prosite, Swiss-Model, ClustalW2, DNAman, DNAstar and MEGA5. [Result] The cloned PpCCR (P. purpureum CCR) cDNA sequence was 1 316 bp, including a 1 110 bp ORF and 206 bp 3’-UTR. The cloned DNA sequence from PpCCR was 6 133 bp in full-length, containing five exons and four introns. Bioinformatic analysis indicated that PpCCR encoded a polypeptide of 369 amino acids, the secondary structure of which was primarily composed of random coil and α-helix, belonging to NAD-dependent epimerase/dehydratase family, and its co-factor binding sites and substrate binding sites were highly conserved. [Conclusion] DNA and cDNA sequences of CCR gene were obtained from P. purpureum, which had the typical characteristics of other homologous genes. The obtained bioinformatic data provided theoretical references for the further analysis of CCR and better application of P. purpureum in the future.

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期刊信息
  • 《农业科学与技术:英文版》
  • 主管单位:湖南省农业科学院
  • 主办单位:湖南省农业科学院
  • 主编:袁隆平
  • 地址:湖南省长沙市芙蓉区马坡岭远大二路892号省农科院内
  • 邮编:410125
  • 邮箱:agricultural2008@163.com
  • 电话:0731-84691326
  • 国际标准刊号:ISSN:1009-4229
  • 国内统一刊号:ISSN:43-1422/S
  • 邮发代号:
  • 获奖情况:
  • 国内外数据库收录:
  • 美国化学文摘(网络版),英国农业与生物科学研究中心文摘,美国剑桥科学文摘
  • 被引量:1218