铁棍山药<italic style="font-style: italic">SUS</italic>基因CDS区克隆与生物信息学分析

刘苏伟; 文艺; 张骆琪; 高素霞; 刘玉霞; 王飞; 鲁传涛; 刘红彦

doi:10.13422/j.cnki.syfjx.20190917

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铁棍山药SUS基因CDS区克隆与生物信息学分析

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- 铁棍山药SUS基因CDS区克隆与生物信息学分析
- Cloning and Bioinformatics Analysis on CDS of SUS Gene in Dioscorea opposita
- 中国实验方剂学杂志 2019年25卷第9期页码：136-143
- 作者机构：
  
  1.河南中医药大学　药学院，郑州　 450046
  2.河南省农业科学院　植物保护研究所，郑州　 450002
- 作者简介：
  
  刘苏伟，在读硕士，从事中药材规范化种植研究，E-mail：2207346588@qq.com
  鲁传涛，博士，研究员，从事中药材病虫害防治研究，Tel：0371-65717380，E-mail：chuantaolu@qq.com
  刘红彦，博士生导师，研究员，从事中药材规范化种植研究，Tel：0371-65730166，E-mail：liμhy1219@163.com；
- 基金信息：
  
  国家中药材产业技术体系郑州综合实验站(CARS-21-12);河南省科技攻关计划项目(162102310054)
- DOI：10.13422/j.cnki.syfjx.20190917
  中图分类号： R284.1; R289; R22; R2-031
- 收稿日期：2018-11-26，
  
  网络出版日期：2019-01-21，
  
  纸质出版日期：2019-05-05
- 稿件说明：
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刘苏伟, 文艺, 张骆琪, 等. 铁棍山药SUS基因CDS区克隆与生物信息学分析[J]. 中国实验方剂学杂志, 2019,25(9):136-143.

Su-wei LIU, Yi WEN, Luo-qi ZHANG, et al. Cloning and Bioinformatics Analysis on CDS of SUS Gene in Dioscorea opposita[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(9): 136-143.
刘苏伟, 文艺, 张骆琪, 等. 铁棍山药SUS基因CDS区克隆与生物信息学分析[J]. 中国实验方剂学杂志, 2019,25(9):136-143. DOI： 10.13422/j.cnki.syfjx.20190917.

Su-wei LIU, Yi WEN, Luo-qi ZHANG, et al. Cloning and Bioinformatics Analysis on CDS of SUS Gene in Dioscorea opposita[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(9): 136-143. DOI： 10.13422/j.cnki.syfjx.20190917.

摘要

目的：

对铁棍山药蔗糖合成酶(sucrose synthase，

SUS

)基因的编码区域(coding sequence

CDS)进行克隆和蛋白质结构分析，为该基因的调控机制与山药多糖的合成机制提供理论依据。

方法：

提取铁棍山药总RNA并反转录为cDNA第一链，根据本实验室铁棍山药基因组数据经注释得到的

SUS

基因序列设计一对特异性引物，利用基因克隆技术获得

SUS

基因的编码区域并通过蛋白质预测分析软件分析蛋白质序列特征。

结果：

克隆得到一个长度2 448 bp的基因序列，具有一个完整的开放阅读框架(open reading frame

ORF)，命名为

DoSUS

1。

DoSUS

1的分子式为C

4209

6534

1115

1205

，相对分子质量9 788.32，共815个氨基酸，理论等电点(PI)6.10，消光系数为110 505，脂溶性指数(AI)为94.15，不稳定指数为32.18，平均亲水指数(GRAVY)为－0.225，属于稳定可溶性酸性蛋白质。

DoSUS

1氨基酸序列存在多个磷酸化位点，不存在跨膜区与信号肽。蛋白质二级结构与三级结构结果显示

DoSUS

1属于全

类蛋白质。功能域预测结果显示

DoSUS

1有蔗糖合成与糖基转移两个功能域。同源性比对结果显示

DoSUS

1的氨基酸序列与所比对单子叶植物的氨基酸序列相似性均

80%。系统进化树显示

DoSUS

1与单子叶植物的SUS进化关系较近。

结论：

首次从铁棍山药中克隆出

SUS

基因的编码序列，并对其蛋白质结构进行了分析，为进一步阐明

SUS

在山药生长发育和多糖合成机制中的作用奠定了基础。

Abstract

Objective:

To clone CDS sequence of

Dioscorea opposita SUS

gene and analyze the protein structure

in order to provide a theoretical basis for the regulation mechanism of

SUS

gene and the synthesis mechanism of

opposita

polysaccharides.

Method:

Total RNA in

opposita

was extracted and reverse-transcribed into first strand of cDNA. Specific primers were designed according to an annotated

SUS

gene sequence obtained from the laboratory

opposita

genome database

and the coding region of the

SUS

gene was obtained by gene cloning technique and the protein sequence characteristics were analyzed by protein prediction analysis software.

Result:

A 2 448 bp gene sequence was cloned with a complete open reading frame (ORF). The gene was named

DoSUS

1.The formula of protein encoded by

DoSUS

1 gene in

opposita

was C

4209

6534

1115

1205

and the molecular weight was 9 788.32

the total number of amino acids was 815

the theory isoelectric point(PI) was 6.10

the extinction coefficient was 110 505

the aliphatic index(AI) was 94.15

the instability index was 32.18

and the grand average of hydropathicity(GRAVY) was -0.225.It was a stable and soluble acidic protein. There were phosphorylation sites in the

DoSUS

1 amino acid sequence

with no transmembrane region and signal peptide. The secondary structure and the tertiary structure showed that

DoSUS

1 was an

class protein. Functional domain predictions showed that

DoSUS

1 had sucrose synthesis domain and glycosyl transfer domain. The homology alignment showed that the amino acid sequence of

DoSUS

1 was more than 80% similar to the amino acid sequence of the aligned monocots. The phylogenetic tree showed that

DoSUS

1 was closely related to SUS of monocotyledon evolution.

Conclusion:

The coding sequence of

SUS

gene was cloned from

opposita

for the first time

and its protein structure was analyzed to lay a foundation for further studying the roles of SUS in the growth and polysaccharide synthesis of

opposita

关键词

Keywords

references

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铁棍山药SUS基因CDS区克隆与生物信息学分析

Cloning and Bioinformatics Analysis on CDS of SUS Gene in Dioscorea opposita

DOI：10.13422/j.cnki.syfjx.20190917

摘要

Abstract

关键词

Keywords

references