Characterization and expression of codon optimized soybean phytase gene in E. coli.

Simple item page
dc.contributor.authorSingh, Pritee
dc.contributor.authorPunjabi, Mansi
dc.contributor.authorJolly, Monica
dc.contributor.authorRai, R D
dc.contributor.authorSachdev, Archana
dc.date.accessioned2014-12-26T04:44:29Z
dc.date.available2014-12-26T04:44:29Z
dc.date.issued2013-12
dc.description.abstractPhytic acid, the major storage form of phosphorus in plant seeds is degraded by the phytases to yield inositol and free phosphate, contributing thereby to the improved bioavailability of phytate phosphorus and essential minerals in plant foods and simultaneous reduction in phosphorus pollution of the terrestrial and aquatic ecosystems. As a possible strategy for altering seed phytate levels, the approach involving reduction of phytate content by ectopically expressing endogenous phytase gene during seed development of soybean (Glycine max L. cv. Pusa-20) was attempted in the present study. Semi-quantitative RT-PCR revealed the maximum expression of phytase gene transcripts in germinating cotyledons (~10 days after germinations), compared to other vegetative tissues. A full-length phytase cDNA was amplified from the germinating seedlings by splicing by overlap extension (SOE)-PCR and its sequence analysis revealed an open-reading-frame of 1644 bp, including an N terminal signal peptide of 28 amino acids. Predicted amino acid sequence (547-aa) of molecular mass 62 kDa on alignment with related purple acid phosphatases in other plants shared five conserved domains and seven invariant amino acids involved in coordination of the metals in the binuclear center of purple acid phosphatases. Owing to a large number of E. coli low-usage codons in soybean phytase gene, the modified gene was cloned into a prokaryotic expression vector pET-28a (+) and its expression in E. coli was confirmed by SDS-PAGE and Western blot analysis. Bioassay of the crude expression product in E. coli revealed a functional phytase gene, showing a great potential for developing low phytate transgenic soybean through its seed-specific overexpression in the early stages of seed development.en_US
dc.identifier.citationSingh Pritee, Punjabi Mansi, Jolly Monica, Rai R D, Sachdev Archana. Characterization and expression of codon optimized soybean phytase gene in E. coli. Indian Journal of Biochemistry & Biophysics. 2013 Dec; 50(6): 537-547.en_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/154210
dc.language.isoenen_US
dc.source.urihttps://nopr.niscair.res.in/handle/123456789/25169en_US
dc.subjectGlycine maxen_US
dc.subjectPhytase geneen_US
dc.subjectSOE-PCRen_US
dc.subjectProkaryotic expressionen_US
dc.subjectE. colien_US
dc.subject.mesh6-Phytase --biosynthesis
dc.subject.mesh6-Phytase --chemistry
dc.subject.mesh6-Phytase --genetics
dc.subject.meshAmino Acid Sequence
dc.subject.meshCloning, Molecular
dc.subject.meshCodon --genetics
dc.subject.meshDNA, Complementary --genetics
dc.subject.meshEscherichia coli --genetics
dc.subject.meshGene Expression
dc.subject.meshGene Expression Regulation, Plant
dc.subject.meshGenetic Engineering --methods
dc.subject.meshMinerals --metabolism
dc.subject.meshMolecular Sequence Data
dc.subject.meshOrgan Specificity
dc.subject.meshPhosphorus --metabolism
dc.subject.meshPhylogeny
dc.subject.meshSeedling --genetics
dc.subject.meshSequence Homology
dc.subject.meshSoybeans --enzymology
dc.subject.meshSoybeans --genetics
dc.subject.meshSoybeans --metabolism
dc.titleCharacterization and expression of codon optimized soybean phytase gene in E. coli.en_US
dc.typeArticleen_US
Files
Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
ijbb2013v50n6p537.pdf
Size:
1.89 MB
Format:
Adobe Portable Document Format
Description:
Jouranl artical
Download
License bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
license.txt
Size:
1.71 KB
Format:
Item-specific license agreed upon to submission
Description:
Download
Collections
Indian Journal of Biochemistry & Biophysics