大叶蛇葡萄类纤维素合成酶基因的克隆及其序列分析

蒲天珍; 张秀桥; 周佩娜; 熊晓妹; 龚玲

doi:10.13422/j.cnki.syfjx.20192012

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大叶蛇葡萄类纤维素合成酶基因的克隆及其序列分析

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- 大叶蛇葡萄类纤维素合成酶基因的克隆及其序列分析
- Cloning and Sequence Analysis of Cellulose Synthase-like Gene from Ampelopsis megalophylla
- 中国实验方剂学杂志 2019年25卷第20期页码：147-152
- 作者机构：
  
  湖北中医药大学　药学院，武汉　 430065
- 作者简介：
  
  蒲天珍，在读硕士，从事中药品种鉴定及新资源开发研究，E-mail：2660384868@qq.com
  龚玲，博士，讲师，从事中药品种鉴定及新资源开发研究，Tel：027-68890106，E-mail：g11224@163.com
- 基金信息：
  
  湖北省教育厅中青年人才项目(Q20182003)
- DOI：10.13422/j.cnki.syfjx.20192012
  中图分类号： R289; R284.1; R22; R2-031
- 收稿日期：2019-01-16，
  
  网络出版日期：2019-07-05，
  
  纸质出版日期：2019-10-20
- 稿件说明：
移动端阅览
蒲天珍, 张秀桥, 周佩娜, 等. 大叶蛇葡萄类纤维素合成酶基因的克隆及其序列分析[J]. 中国实验方剂学杂志, 2019,25(20):147-152.

Tian-zhen PU, Xiu-qiao ZHANG, Pei-na ZHOU, et al. Cloning and Sequence Analysis of Cellulose Synthase-like Gene from Ampelopsis megalophylla[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(20): 147-152.
蒲天珍, 张秀桥, 周佩娜, 等. 大叶蛇葡萄类纤维素合成酶基因的克隆及其序列分析[J]. 中国实验方剂学杂志, 2019,25(20):147-152. DOI： 10.13422/j.cnki.syfjx.20192012.

Tian-zhen PU, Xiu-qiao ZHANG, Pei-na ZHOU, et al. Cloning and Sequence Analysis of Cellulose Synthase-like Gene from Ampelopsis megalophylla[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(20): 147-152. DOI： 10.13422/j.cnki.syfjx.20192012.

摘要

目的：

从药用植物大叶蛇葡萄

Ampelopsis megalophylla

中克隆得到类纤维素合酶(Cellulose synthase-like，Csl)基因(

AmCsl

)全长，并对序列进行生物信息学分析。

方法：

根据大叶蛇葡萄

megalophylla

转录组测序数据得到的

AmCsl

基因序列设计特异引物，以嫩叶总RNA逆转录的cDNA为模板，采用聚合酶链式反应(PCR)技术扩增

AmCsl

基因全长，并进行TA克隆测序，利用多种软件对该序列进行生物信息学分析。

结果：

AmCsl

全长cDNA为1 438 bp，包含1个561 bp的完整开放阅读框(open reading frame，ORF)，编码186个氨基酸，蛋白质分子式为C

1011

1547

233

257

，理论相对分子质量为22.40 kDa，理论等电点(PI)为7.59，脂肪族指数(AI)为116.88，存在跨膜区，无信号肽，可能定位于内质网膜，平均疏水系数为0.670，不稳定指数为42.56，属于疏水性不稳定蛋白，保守结构域包含了1个纤维素合成酶超家族结构域，二级结构以

-螺旋为主。氨基酸多序列比对和系统进化树分析发现，

AmCsl

与同科植物葡萄有较高的同源性。

结论：

获得了大叶蛇葡萄

AmCsl

基因的全长，对其进行了初步的生物信息学分析，为进一步研究大叶蛇葡萄多糖的积累和生物合成的调控奠定了必要基础。

Abstract

Objective:

To clone cellulose synthase-like(Csl)gene from ethnic medicinal plant

Ampelopsis megalophylla

and analyze its sequence by bioinformatics.

Method:

Specific primers were designed for

AmCsl

gene sequences obtained from

megalophylla

transcriptome sequencing data. The full-length cDNA of

AmCsl

gene was amplified by PCR using cDNA of young leaves as template

and TA clone and sequencing was performed. The sequence was analyzed by bioinformatics.

Result:

The full length cDNA was 1 438 bp

containing a 561 bp open reading frame(ORF)

and encoding 186 amino acids

the molecular formula of protein was C

1011

1547

233

257

the theoretical relative molecular weight was 22.40 kDa

the theory isoelectric point(PI)was 7.59

and the aliphatic index(AI)was 116.88.There was a transmembrane region and no signal peptide

which may be located in the endoplasmic reticulum

the average hydrophobic coefficient was 0.670

and the instability index was 42.56.It belonged to a hydrophobic unstable protein. The conserved domain contained a cellulose synthase

and the secondary structure mainly was dominated by

-helix. Amino acid sequence alignment and phylogenetic tree analysis showed that

AmCsl

had a high homology with

Vitis vinifera

Conclusion:

The full length of

AmCsl

gene was obtained for preliminary bioinformatics analysis

which laid a necessary foundation for further study on the accumulation of polysaccharides and the regulation of biosynthesis.

关键词

Keywords

references

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