Abstract
Thaumatin-like proteins (TLPs) constitutes a homogeneous family, members of which are produced by plants in response to different kinds of stress. NP24 protein is one of such salt-induced protein from tomato (Solanum lycopersicum) and it belongs to TLPs family. NP24 is a 24 kDa (207 amino acid) antimicrobial TLP found in tomato fruits. One isoform (NP24I) of NP24 was discovered in the outer pericarp of tomato fruit and is reported to play a possible role in ripening of the fruit in addition to its antimicrobial activity. In this study, the total RNA was isolated successfully from the outer pericarp of ripe (red) tomato fruit. cDNA was prepared and the gene coding for NP24I protein was amplified using conventional polymerase chain reaction (PCR). The gene was then cloned into Mach1™- T1® Escherichia colicells, then subcloned into the over-expression vector pET-28a (+) using BL21expression bacteria. Sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) analysis showed that the gene was over-expressed in E. coli as inclusion bodies. Optimization of the recombinant NP24I protein solubility was achieved by cold induction through decreasing both expression temperature andIsopropyl-beta-thio galactopyranoside (IPTG) concentration. The recombinant NP24I protein was purified using Ni-NTA resin, and then the antimicrobial activity of the purified recombinant NP24I protein was tested. The aims of this work were to study the cloning and expression of NP24 protein from local tomato cultivar in a prokaryotic system and to test the activity of the recombinant NP24, as well as to prove the activity of native protein on the bacterial as well as fungal growth. Key words: Pathogenesis-related proteins (PR), low molecular weight thaumatin-like protein (TLPs), cloning, Escherichia coli, cold induction, antimicrobial activity, tomato (Solanum lycopersicum).
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Claims 5
Claim · model-written summary
NP24I was identified in the outer pericarp of tomato fruit.
- Taxon
- tomato
Solanum lycopersicume0 - Organ / stage
- outer pericarp of tomato fruite0
- Gene
- NP24Ie0
- Outcome
- discovered in the outer pericarpe0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“One isoform (NP24I) of NP24 was discovered in the outer pericarp of tomato fruit”
Claim · model-written summary
The NP24I gene was amplified from tomato fruit cDNA.
- Taxon
- tomatoSolanum lycopersicumt0
- Gene
- NP24I proteine0
- Outcome
- gene coding for NP24I protein was amplifiede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“cDNA was prepared and the gene coding for NP24I protein was amplified using conventional polymerase chain reaction (PCR)”
t0“tomato fruit”
Claim · model-written summary
NP24I was overexpressed as inclusion bodies in E. coli.
- Taxon
- tomatoSolanum lycopersicumt0
- Gene
- genee0
- Outcome
- over-expressed in E. coli as inclusion bodiese0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the gene was over-expressed in E. coli as inclusion bodies”
t0“tomato fruit”
Claim · model-written summary
Cold induction optimized recombinant NP24I solubility by lowering temperature and IPTG concentration.
- Taxon
- tomatoSolanum lycopersicumt0
- Treatment
- cold inductione0
- Gene
- NP24I proteine0
- Outcome
- recombinant NP24I protein solubilitye0
- Direction
- Optimizatione0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Optimization of the recombinant NP24I protein solubility was achieved by cold induction through decreasing both expression temperature andIsopropyl-beta-thio galactopyranoside (IPTG) concentration”
t0“tomato fruit”
Claim · model-written summary
Recombinant NP24I protein was purified with Ni-NTA resin.
- Taxon
- tomatoSolanum lycopersicumt0
- Gene
- NP24I proteine0
- Outcome
- purified using Ni-NTA resine0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“recombinant NP24I protein was purified using Ni-NTA resin”
t0“tomato fruit”
The same questions in other papers
cold · quality: protein · Solanum lycopersicum (tomato)Positive and negative results
1 paper in the same direction as this paper · 1 in the opposite direction
- negative
- positive
Claims are grouped by organism, treatment and what was measured or targeted. A paper counts as positive when any of its claims in the group is positive.