Record Details

Characterization of squalene synthase gene from Gymnema sylvestre R. Br.

KRISHI: Publication and Data Inventory Repository

View Archive Info
 
 
Field Value
 
Title Characterization of squalene synthase gene from Gymnema sylvestre R. Br.
Not Available
 
Creator KULDEEPSINGH KALARIYA
RAM PRASNNA MEENA
LIPI POOJARA
DEEPA SHAHI
Sandip Patel
 
Subject Gymnema sylvestre
Intron
Long-range PCR
Next-generation sequencing
Squalene synthase
 
Description Background: Squalene synthase (SQS) is a rate-limiting enzyme necessary to produce pentacyclic triterpenes in
plants. It is an important enzyme producing squalene molecules required to run steroidal and triterpenoid
biosynthesis pathways working in competitive inhibition mode. Reports are available on information pertaining to
SQS gene in several plants, but detailed information on SQS gene in Gymnema sylvestre R. Br. is not available. G.
sylvestre is a priceless rare vine of central eco-region known for its medicinally important triterpenoids. Our work
aims to characterize the GS-SQS gene in this high-value medicinal plant.
Results: Coding DNA sequences (CDS) with 1245 bp length representing GS-SQS gene predicted from
transcriptome data in G. sylvestre was used for further characterization. The SWISS protein structure modeled for the
GS-SQS amino acid sequence data had MolProbity Score of 1.44 and the Clash Score 3.86. The quality estimates
and statistical score of Ramachandran plots analysis indicated that the homology model was reliable. For full-length
amplification of the gene, primers designed from flanking regions of CDS encoding GS-SQS were used to get
amplification against genomic DNA as template which resulted in approximately 6.2-kb sized single-band product.
The sequencing of this product through NGS was carried out generating 2.32 Gb data and 3347 number of
scaffolds with N50 value of 457 bp. These scaffolds were compared to identify similarity with other SQS genes as
well as the GS-SQSs of the transcriptome. Scaffold_3347 representing the GS-SQS gene harbored two introns of 101
and 164 bp size. Both these intronic regions were validated by primers designed from adjoining outside regions of
the introns on the scaffold representing GS-SQS gene. The amplification took place when the template was
genomic DNA and failed when the template was cDNA confirmed the presence of two introns in GS-SQS gene in
Gymnema sylvestre R. Br.
Conclusion: This study shows GS-SQS gene was very closely related to Coffea arabica and Gardenia jasminoides and
this gene harbored two introns of 101 and 164 bp size.
 
Date 2021-01-30T11:25:36Z
2021-01-30T11:25:36Z
2021-01-16
 
Type Research Paper
 
Identifier Kalariya, K.A., Meena, R.P., Poojara, L. et al. Characterization of squalene synthase gene from Gymnema sylvestre R. Br.. Beni-Suef Univ J Basic Appl Sci 10, 6 (2021). https://doi.org/10.1186/s43088-020-00094-4
Not Available
http://krishi.icar.gov.in/jspui/handle/123456789/45161
 
Language English
 
Relation Not Available;
 
Publisher SPRINGER OPEN