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1.北京中医药大学 生命科学学院,北京 102488
2.宁夏医科大学 药学院,银川 750004
[第一作者] 侯嘉铭,在读硕士,从事药用植物功能基因与分子生药学研究,E-mail:18829034555@163.com
*王汉卿,博士,副教授,从事中药资源研究,E-mail:wwwhhq@163.com
*刘颖,博士,副教授,从事药用植物功能基因与分子生药学研究,Tel:010-84738646,E-mail:liuyliwd@sina.com;
收稿日期:2019-04-29,
网络出版日期:2019-10-11,
纸质出版日期:2020-03-20
移动端阅览
侯嘉铭, 田少凯, 杨林, 等. 光果甘草
Jia-ming HOU, Shao-kai TIAN, Lin YANG, et al. Molecular Cloning and Sequence Analysis of
侯嘉铭, 田少凯, 杨林, 等. 光果甘草
Jia-ming HOU, Shao-kai TIAN, Lin YANG, et al. Molecular Cloning and Sequence Analysis of
目的:
2
通过克隆光果甘草
Glycyrrhiza glabra
UDP-葡萄糖4-差向异构酶(UDP-glucose 4-epimerase,
UGE
)基因并进行生物信息学分析,探究光果甘草
UGE
基因与甘草酸生物合成分子调控之间的潜在关系。
方法:
2
从光果甘草主根中提取RNA,通过逆转录聚合酶链式反应(RT-PCR)克隆得到
UGE
基因cDNA序列,测序并进行序列分析。
结果:
2
成功克隆得到一条长度为1 121 bp的光果甘草
UGE
基因(
GgUGE
)cDNA序列,包含1个长度为1 053 bp的开放阅读框(ORF),共编码350个氨基酸残基,在GenBank上对所得cDNA序列进行注册,序列注册号为MK638908。序列分析表明
GgUGE
基因编码蛋白为不稳定亲水蛋白,理论相对分子质量为39.02 kDa,等电点为6.13,不含信号肽,无跨膜区,二级结构以
α
-螺旋为主,保守结构域含有葡萄糖4-差向异构酶基因家族结构域。
GgUGE
基因的cDNA序列和氨基酸序列聚类分析结果表明其与豆科植物亲缘关系最近,而与杨柳科植物亲缘关系较远。
结论:
2
首次克隆得到了光果甘草
GgUGE
cDNA序列,对其进行了生物信息学分析,为后续
GgUGE
基因的功能研究以及甘草酸生物合成分子调控机制的解析提供参考。
Objective:
2
To clone the cDNA sequence of UDP-glucose 4-epimerase (UGE) in
Glycyrrhiza glabra
and analyze its sequence
so as to explore the potential relationship between the
UGE
gene and the molecular regulatory mechanisms of glycyrrhizic acid biosynthesis.
Method:
2
The cDNA sequence of
UGE
was cloned from the root of
G
.
glabra
by reverse transcription polymerase chain reaction (RT-PCR)
then sequenced and analyzed by bioinformatics software.
Results:
2
A
GgUGE
cDNA sequence with the full length of 1 121 bp was obtained. The open reading fame (ORF) of
GgUGE
was 1 053 bp
encoding 350 amino acid residues. The
GgUGE
cDNA sequence was submitted to GenBank
and the accession No. was MK638908. Sequence analysis showed that GgUGE was an unstable hydrophilic protein
its average relative molecular weight was 39.02 kDa
and isoelectric point was 6.13. It contained no signal peptides or transmembrane domains. Its secondary structure mainly constituted of
α
-helix and had a conversed domain of UDP-glucose 4-epimerase superfamily. The homologoue analysis showed that the cDNA and amino acid sequences of
GgUGE
had the closest evolutionary relationship to Leguminosae and relatively distant evolutionary relationship to Salicaceae.
Conclusion:
2
In this study
GgUGE cDNA
sequence is successfully cloned from
G
.
glabra
for the very first time
which will provide reference for studying the function of
GgUGE
and explaining the molecular regulatory mechanisms of glycyrrhizic acid biosynthesis in
G
.
glabra
.
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