红花DHDPS基因全长cDNA的克隆及植物表达载体构建

发布时间：2018-05-03 06:21

本文选题：红花 + 二氢吡啶二羧酸合酶　；参考：《中草药》2017年08期

【摘要】：目的克隆红花赖氨酸生物合成途径关键酶二氢吡啶二羧酸合酶(dihydrodipicolinate synthase DHDPS)基因的全长c DNA序列,构建植物超表达载体。方法根据红花转录组文库注释信息获得红花DHDPS(CtDHDPS)基因的中间序列,通过RT-PCR与RACE技术从红花种子中克隆CtDHDPS基因全长c DNA序列。采用DNA重组技术构建p BASTA-CtDHDPS植物超表达载体。结果分离CtDHDPS基因全长1 309 bp,开放阅读框954 bp,编码317个氨基酸,编码蛋白理论等电点为5.93,相对分子质量约为34 750.79。通过DNA重组技术成功构建了植物超表达载体p BASTA-CtDHDPS。结论获得CtDHDPS基因全长c DNA序列并成功构建植物超表达载体,为阐明CtDHDPS基因的生物学功能及其在红花赖氨酸生物合成途径中作用机制提供科学依据。
[Abstract]:Objective to clone the full-length c DNA sequence of dihydrodipicolinate synthase synthase, a key enzyme in the biosynthesis pathway of safflower lysine, and to construct a plant overexpression vector. Methods according to the annotated information of Carthamus transcriptome library, the intermediate sequence of CtDHDPS gene was obtained, and the full-length c DNA sequence of CtDHDPS gene was cloned from the seeds of Carthamus safflower by RT-PCR and RACE techniques. The plant superexpression vector of p BASTA-CtDHDPS was constructed by DNA recombination technique. Results the total length of CtDHDPS gene was 1 309 BP, open reading frame 954 BP, encoding 317 amino acids, the theoretical isoelectric point of encoding protein was 5.93, and the relative molecular weight was about 34 750.79. The plant superexpression vector pBASTA-CtDHDPSS was successfully constructed by DNA recombination technique. Conclusion the full-length c DNA sequence of CtDHDPS gene was obtained and the plant overexpression vector was successfully constructed, which provides a scientific basis for elucidating the biological function of CtDHDPS gene and its mechanism in the biosynthesis pathway of lysine in safflower.
【作者单位】：吉林农业大学中药材学院;吉林农业大学生物反应器与药物开发教育部工程研究中心;吉林农业大学科技管理处;吉林农业大学生命科学学院;
【基金】：国家自然科学基金资助项目(31401445) 吉林省教育厅“十三五”科学技术项目(JJKH20170310KJ) 吉林省科技厅医药产业推进计划项目(20140311034YY) 国家高技术研究发展计划(“863”)项目(2011AA100606) 2017年吉林农业大学大学生创新创业训练计划项目
【分类号】：S567.219

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