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北京中医药大学 生命科学学院,北京 102488
[第一作者] 田少凯,在读硕士,从事药用植物功能基因与分子生药学研究,E-mail:tiansk0310@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,E-mail:liuyliwd@sina.com
收稿日期:2019-04-29,
网络出版日期:2019-06-19,
纸质出版日期:2020-02-20
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田少凯, 侯嘉铭, 高智强, 等. 光果甘草
Shao-kai TIAN, Jia-ming HOU, Zhi-qiang GAO, et al. Cloning and Bioinformatic Analysis of ARPI Gene from Glycyrrhiza glabra[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(4): 185-190.
田少凯, 侯嘉铭, 高智强, 等. 光果甘草
Shao-kai TIAN, Jia-ming HOU, Zhi-qiang GAO, et al. Cloning and Bioinformatic Analysis of ARPI Gene from Glycyrrhiza glabra[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(4): 185-190. DOI: 10.13422/j.cnki.syfjx.20191951.
目的:
2
克隆光果甘草生长素/吲哚乙酸蛋白(Aux/IAA)(GgARPI)基因并进行生物信息学分析。
方法:
2
取光果甘草新鲜主根,提取RNA,通过逆转录-聚合酶链式反应(RT-PCR)克隆
GgARPI
基因互补脱氧核糖核酸(cDNA)序列,测序并对其进行生物信息学分析。
结果:
2
克隆得到长度为686 bp的
GgARPI
基因cDNA序列,包含1个585 bp的完整开放阅读框(ORF),编码194个氨基酸残基。生物信息学分析表明
GgARPI
编码蛋白为稳定亲水蛋白,相对分子质量21.95 kDa,等电点6.85,不含信号肽,无跨膜区,二级结构以无规卷曲为主,包含1个Aux/IAA蛋白超家族结构域。同源性分析表明
GgARPI
基因与豆科植物的亲缘关系较近,与单子叶植物谷子
Setaria italica
的进化关系最远。
结论:
2
首次克隆得到
GgARPI
cDNA序列,为进一步研究
GgARPI
基因的功能及其对甘草酸生物合成的分子调控机制奠定了基础。
Objective:
2
To clone the complementary deoxyribonucleic acid (cDNA) of auxin/indole acetic acid protein (Aux/IAA) from
Glycyrrhiza glabra
(GgARPI) and analyze its sequence by bioinformatics.
Method:
2
RNA was extracted from fresh root of
G
.
glabra
the cDNA sequence of
GgARPI
gene was cloned by reverse transcription polymerase chain reaction (RT-PCR)
then sequencing and bioinformatic analysis were performed.
Result:
2
The
GgARPI
cDNA sequence with the full length of 686 bp was obtained from
G
.
glabra
. The full open reading frame (ORF) was 585 bp
encoding 194 amino acid residues. The bioinformatic analysis showed that the protein coded by
GgARPI
was a stable hydrophilic protein
with a relative molecular weight of 21.95 kDa and isoelectric point of 6.85.It contained no signal peptides or transmembrane domain. Its secondary structure mainly consisted of random coil. An Aux/IAA superfamily was included in the conversed domain. Homology analysis indicated that it had a close evolutionary relationship with leguminous plants
and a distant evolutionary relationship with monocotyledon
such as
Setaria italica
.
Conclusion:
2
GgARPI
cDNA sequence is successfully cloned from
G
.
glabra
for the first time
which will lay a foundation for studying the function of
GgARPI
and explaining the molecular regulatory mechanism of biosynthesis of glycyrrhizic acid in
G
.
glabra
.
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