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Molecular Cloning and Characterization of Hydroperoxide Lyase Gene in the Leaves of Tea Plant (Camellia sinensis)
- Source :
- Journal of Agricultural and Food Chemistry. 64:1770-1776
- Publication Year :
- 2016
- Publisher :
- American Chemical Society (ACS), 2016.
-
Abstract
- Hydroperoxide lyase (HPL, E.C. 4.1.2.) is the major enzyme in the biosynthesis of natural volatile aldehydes and alcohols in plants, however, little was known about HPL in tea plants (Camellia sinensis). A unique cDNA fragment was isolated by suppressive subtractive hybridization (SSH) from a tea plant subjected to herbivory by tea geometrid Ectropis obliqua. This full length cDNA acquired by RACE was 1476 bp and encoded 491 amino acids. DNA and protein BLAST searches showed high homology to HPL sequences from other plants. The His-tag expression vector pET-32a(+)/CsHPL was constructed and transferred into Escherichia coli Rosetta (DE3). The expression product of recombinant CsHPL in E. coli was about 60 kDa. The enzyme activity of CsHPL was 0.20 μmol·min(-1)·mg(-1). Quantitative RT-PCR analysis indicated CsHPL was strongly up-regulated in tea plants after Ectropis obliqua attack, suggesting that it may be an important candidate for defense against insects in tea plants.
- Subjects :
- 0106 biological sciences
0301 basic medicine
Molecular Sequence Data
Moths
Molecular cloning
medicine.disease_cause
01 natural sciences
Camellia sinensis
03 medical and health sciences
Cytochrome P-450 Enzyme System
Gene Expression Regulation, Plant
Complementary DNA
medicine
Animals
Amino Acid Sequence
Cloning, Molecular
Escherichia coli
Aldehyde-Lyases
Plant Proteins
Expression vector
biology
food and beverages
General Chemistry
Molecular biology
Enzyme assay
Plant Leaves
030104 developmental biology
Biochemistry
Ectropis obliqua
Suppression subtractive hybridization
biology.protein
General Agricultural and Biological Sciences
Sequence Alignment
010606 plant biology & botany
Subjects
Details
- ISSN :
- 15205118 and 00218561
- Volume :
- 64
- Database :
- OpenAIRE
- Journal :
- Journal of Agricultural and Food Chemistry
- Accession number :
- edsair.doi.dedup.....868e9a3c24da16d92ce4522e3de9be92