Evidence map›Paper›PMID 34845523›Full record

ArticlePlanta2021

Molecular cloning and functional characterization of two squalene synthase genes in Atractylodes lancea.

Junxian Wu, Rui Xu, Jimei Lu, Weiwei Liu, Hanwen Yu, Mengli Liu, Jing Li, Minzhen Yin, Huasheng Peng, Liangping Zha

Abstract read
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In one paragraph

Article in Planta, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.

0numbers the graph read from it
0cells of the map it votes in
4citing papers in PubMed
0.7field-weighted citation impact, top 30% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

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3 · Its place in the literature

Who cites it

4 citing papers in PubMed, 11 citations in OpenAlex.

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4 · The record

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PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

10 authors at 2 institutions in 1 country.

Junxian Wu *College of Pharmacy, Anhui University of Chinese Medicine, Hefei, 230012, China.
Rui Xu *College of Pharmacy, Anhui University of Chinese Medicine, Hefei, 230012, China.
Jimei LuCollege of Pharmacy, Anhui University of Chinese Medicine, Hefei, 230012, China.
Weiwei LiuCollege of Pharmacy, Anhui University of Chinese Medicine, Hefei, 230012, China.
Hanwen YuCollege of Pharmacy, Anhui University of Chinese Medicine, Hefei, 230012, China.
Mengli LiuCollege of Pharmacy, Anhui University of Chinese Medicine, Hefei, 230012, China.
Jing LiCollege of Pharmacy, Anhui University of Chinese Medicine, Hefei, 230012, China.
Minzhen YinCollege of Pharmacy, Anhui University of Chinese Medicine, Hefei, 230012, China.
Huasheng PengCollege of Pharmacy, Anhui University of Chinese Medicine, Hefei, 230012, China. hspeng@126.com.
Liangping ZhaCollege of Pharmacy, Anhui University of Chinese Medicine, Hefei, 230012, China. zlp_ahtcm@126.com.ORCID http://orcid.org/0000-0002-4644-5852
Anhui University of Traditional Chinese Medicine · CNChinese Academy of Medical Sciences & Peking Union Medical College · CN

Funding

National Natural Science Foundation of China 82073957
6 · The paper itself

Abstract

MAIN

conclusionTwo squalene synthase genes AlSQS1 and AlSQS2 were isolated from Atractylodes lancea and functionally characterized using in vitro enzymatic reactions. Atractylodes lancea is a traditional herb used for the treatment of rheumatic diseases, gastric disorders, and influenza. Its major active ingredients include sesquiterpenoids and triterpenes. Squalene synthase (SQS; EC 2.5.1.21) catalyzes the first enzymatic step in the central isoprenoid pathway towards sterol and triterpenoid biosynthesis. In this study, we aimed to investigate two SQSs from A. lancea using cloning and in vitro enzymatic characterization. Bioinformatics and phylogenetic analyses revealed that the AlSQSs exhibited high homology with other plant SQSs. Furthermore, AlSQS1 was observed to be localized in both the nucleus and cytoplasm, whereas AlSQS2 was localized in the cytoplasm and endoplasmic reticulum. To obtain soluble recombinant enzymes, AlSQS1 and AlSQS2 were successfully expressed as glutathione S-transferase (GST)-tagged fusion proteins in Escherichia coli Transetta (DE3). Approximately 68 kDa recombinant proteins were obtained using GST-tag affinity chromatography and Western blot analysis. Results of the in vitro enzymatic reactions established that both AlSQS1 and AlSQS2 were functional, which verifies their catalytic ability in converting two farnesyl pyrophosphates to squalene. The expression patterns of AlSQS and selected terpenoid genes were also investigated in two A. lancea chemotypes using available RNA sequencing data. AlSQS1 and AlSQS2, which showed relatively similar expression in the three tissues, were more highly expressed in the stems than in the leaves and rhizomes. Methyl jasmonate (MeJA) was used as an elicitor to analyze the expression profiles of AlSQSs. The results of qRT-PCR analysis revealed that the gene expression of AlSQS1 and AlSQS2 plummeted at lowest value at 12 h and reached its peak at 24 h. This study is the first report on the cloning, characterization, and expression of SQSs in A. lancea. Therefore, our findings contribute novel insights that may be useful for future studies regarding terpenoid biosynthesis in A. lancea.

Indexed as

AtractylodesFarnesyl-Diphosphate FarnesyltransferaseCloning, MolecularGenes, PlantPhylogenySequence Analysis, RNASqualeneFarnesyl-Diphosphate FarnesyltransferaseSqualeneAtractylodes lanceaBiosynthesis pathwaysFunctional characterizationSqualene synthaseTerpenoids

Identifiers

PMID34845523
OpenAlexW3217325387

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.