Cloning and Bioinformatics Analysis of 88 Gene in

CHEN Shang; WANG Xiu-juan; ZHANG Yi-feng; GAO Wei

doi:10.13422/j.cnki.syfjx.20180606

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Cloning and Bioinformatics Analysis of 88 Gene in

更新时间：2024-01-04

- Cloning and Bioinformatics Analysis of 88 Gene in
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 6, Pages: 32-36(2018)
- 作者机构：
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- DOI：10.13422/j.cnki.syfjx.20180606
  CLC： S567.19;Q943.2
- Published：2018
- 稿件说明：
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CHEN Shang, WANG Xiu-juan, ZHANG Yi-feng, et al. Cloning and Bioinformatics Analysis of 88 Gene in [J]. Chinese journal of experimental traditional medical formulae, 2018, 24(6): 32-36.
DOI：

CHEN Shang, WANG Xiu-juan, ZHANG Yi-feng, et al. Cloning and Bioinformatics Analysis of 88 Gene in [J]. Chinese journal of experimental traditional medical formulae, 2018, 24(6): 32-36. DOI： 10.13422/j.cnki.syfjx.20180606.

摘要

目的：研究雷公藤萜类成分的生物合成相关基因，在雷公藤转录组信息数据的基础上，以雷公藤悬浮细胞为研究材料，克隆TwCYP88A1基因。方法：采用cDNA末端快速扩增技术（RACE）克隆雷公藤TwCYP88A1基因，实时荧光定量聚合酶链式反应（PCR）分析其组织表达谱。结果： TwCYP88A1 cDNA全长1 594 bp，编码491个氨基酸，等电点9.16，相对分子质量55.957 kDa，具有半胱氨酸铁血红素配体信号活性催化位点。植物组织表达分析表明TwCYP88A1基因在雷公藤叶中的表达量最高，根中最低。结论：从雷公藤悬浮细胞中克隆得到TwCYP88A1基因全长cDNA，对其进行生物信息学分析，揭示其组织表达谱，为深入分析雷公藤细胞色素P450酶、解析雷公藤萜类化合物的生物合成下游途径奠定基础。

Abstract

Objective: To study on the genes related to the biosynthesis of diterpenoid in Tripterygium wilfordii

based on the transcriptome data

TwCYP88A1 gene was cloned from T. wilfordii suspension cells.Method: TwCYP88A1 was obtained by rapid amplification of cDNA ends(RACE).Real-time quantitative polymerase chain reaction (PCR) was used to detect the relative expression profile of TwCYP88A1 in different tissues of T. wilfordii.Result: The full-length cDNA of TwCYP88A1 was 1 594 bp encoding a polypeptide of 491 amino acids.The deduced isoelectric point (pI) was 9.16

a calculated relative molecular weight was about 55.957 kDa.It had active catalytic sites of cysteine hemoglobin ligand signal.Plant tissue expression analysis indicated that TwCYP88A1 expressed the highest in leaves

while lowest in roots.Conclusion: The full-length cDNA of TwCYP88A1 can be cloned from the suspension cells of T. wilfordii.

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