Characterization, cloning, expression and bioassay of vip3 gene isolated from an Egyptian Bacillus thuringiensis against whiteflies

El-Gaied, Lamiaa F.; Mahmoud, Alshimaa; Salem, Reda; Elmenofy, Wael; Saleh, İbrahim; Abulreesh, Hussein H.; Arif, Ibrahim A.; Osman, Gamal

doi:10.1016/j.sjbs.2019.12.013

Cited by 13 publications

(6 citation statements)

References 22 publications

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“…This study demonstrated that the whole-cell culture of Vip3 protein-expressing cells showed more toxicity potential (4.7 times higher LC 50 ) against whiteflies than Vip3 protein alone. Therefore, these proteins can be successfully used in the control and management of whiteflies ( El-Gaied et al, 2020 ). Recently, Vip3Aa19 protein was found to be very effective against A. ipsilon attack in maize with LC 50 value of 0.43 μg/g as compared to chimeric protein Vip3_Ch1 (5.53 μg/g).…”

Section: Vegetative Insecticidal Proteins (Vips)mentioning

confidence: 99%

Vegetative Insecticidal Protein (Vip): A Potential Contender From Bacillus thuringiensis for Efficient Management of Various Detrimental Agricultural Pests

2021

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Bacillus thuringiensis (Bt) bacterium is found in various ecological habitats, and has natural entomo-pesticidal properties, due to the production of crystalline and soluble proteins during different growth phases. In addition to Cry and Cyt proteins, this bacterium also produces Vegetative insecticidal protein (Vip) during its vegetative growth phase, which is considered an excellent toxic candidate because of the difference in sequence homology and receptor sites from Cry proteins. Vip proteins are referred as second-generation insecticidal proteins, which can be used either alone or in complementarity with Cry proteins for the management of various detrimental pests. Among these Vip proteins, Vip1 and Vip2 act as binary toxins and have toxicity toward pests belonging to Hemiptera and Coleoptera orders, whereas the most important Vip3 proteins have insecticidal activity against Lepidopteran pests. These Vip3 proteins are similar to Cry proteins in terms of toxicity potential against susceptible insects. They are reported to be toxic toward pests, which can’t be controlled with Cry proteins. The Vip3 proteins have been successfully pyramided along with Cry proteins in transgenic rice, corn, and cotton to combat resistant pest populations. This review provides detailed information about the history and importance of Vip proteins, their types, structure, newly identified specific receptors, and action mechanism of this specific class of proteins. Various studies conducted on Vip proteins all over the world and the current status have been discussed. This review will give insights into the significance of Vip proteins as alternative promising candidate toxic proteins from Bt for the management of pests in most sustainable manner.

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Section: Vegetative Insecticidal Proteins (Vips)mentioning

confidence: 99%

Vegetative Insecticidal Protein (Vip): A Potential Contender From Bacillus thuringiensis for Efficient Management of Various Detrimental Agricultural Pests

2021

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“…Reasons for using BacSF9 system for protein expression included (1) secretion into the growth medium; (2) production of less aggregated proteins with an increased probability of correct folding, and (3) post-translational modifications for better antigenicity and specificity. Previously, the baculovirus expression system was successfully used to express different viral proteins such as the tomato yellow leaf curl virus coat protein (TYLCV) (El-Gaied, Salem & Elmenofy, 2017), Garlic mite-borne filamentous virus (GarMbFV) (Ardisson-Arau'jo et al, 2013), and glycoprotein E (gE) of the Egyptian BoHV-1.1 Abu-Hammad strain (rBac/gE-AbuH) (El-Kholy et al, 2013;El-Gaied et al, 2020). Furthermore, recombinant proteins produced using this system were validated for their antigenicity.…”

Section: Discussionmentioning

confidence: 99%

Expression of 1B capsid protein of Foot-and-mouth disease virus (FMDV) using baculovirus expression system and its validation in detecting SAT 2- specific antisera

Elmenofy

Mohamed

El-Gaied

et al. 2020

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Foot-and-mouth disease virus (FMDV) is one of the most devastating animal viruses that affect livestock worldwide. The 1B capsid of FMDV has been widely used to detect and confirm the infection. In the present study, the sequence coding for 1B subunit of FMDV capsid was expressed in insect cells using the baculovirus expression system under the polyhedrin (polh) promoter. The expression of 1B capsid protein was validated in the culture filtrate of insect cells using SDS-PAGE and western blotting. The culture filtrate containing recombinant 1B capsid (r1B) was used as a coated antigen in an indirect enzyme-linked immunosorbent assay (ELISA). The antigenicity and specificity of r1B against SAT 2 serotype-specific antibodies were assessed. Our results revealed that a protein concentration as low as 25 ng could detect SAT 2-specific antibodies in ELISA. The results highlight the application of insect cells developed r1B protein in the detection of FMDV. Further studies are required to determine the ability of r1B to detect other FMDV serotypes.

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“…Upon the obtaining sequence, a pair of specific primer was designed (sense: 5′- CGC GGATCC AGCAGCAGCAACTGCGCC-3′, and the anti-sense: 5′- CCG CTCGAG CTAGCAGTTCTTGCAGAAGC-3′ flanked by Bam HI and Xho I recognition sites ( italic ), respectively) for subcloning into pGEX-4 T-1 prokaryotic expression vector. The defensin coding sequence was inserted in-frame to be fused with GST-tagged protein to facilitate the further purification (Salem et al 2019a ; El-Gaied et al 2020 ).…”

Section: Methodsmentioning

confidence: 99%

Utilization of a recombinant defensin from Maize (Zea mays L.) as a potential antimicrobial peptide

et al. 2020

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The search for effective and bioactive antimicrobial molecules to encounter the medical need for new antibiotics is an encouraging area of research. Plant defensins are small cationic, cysteine-rich peptides with a stabilized tertiary structure by disulfide-bridges and characterized by a wide range of biological functions. The heterologous expression of Egyptian maize defensin (MzDef) in Escherichia coli and subsequent purification by glutathione affinity chromatography yielded 2 mg/L of recombinant defensin peptide. The glutathione-S-transferase (GST)-tagged MzDef of approximately 30 kDa in size (26 KDa GST + ~ 4 KDa MzDef peptide) was immunodetected with anti-GST antibodies. The GST-tag was successfully cleaved from the MzDef peptide by thrombin, and the removal was validated by the Tris-Tricine gel electrophoresis. The MzDef induced strong growth inhibition of Rhizoctonia solani, Fusarium verticillioides, and Aspergillus niger by 94.23%, 93.34%, and 86.25%, respectively, whereas relatively weak growth inhibitory activity of 35.42% against Fusarium solani was recorded. Moreover, strong antibacterial activities were demonstrated against E. coli and Bacillus cereus and the moderate activities against Salmonella enterica and Staphylococcus aureus at all tested concentrations (0.1, 0.2, 0.4, 0.8, 1.6, and 3.2 µM). Furthermore, the in vitro MTT assay exhibited promising anticancer activity against all tested cell lines (hepatocellular carcinoma, mammary gland breast cancer, and colorectal carcinoma colon cancer) with IC50 values ranging from 14.85 to 29.85 µg/mL. These results suggest that the recombinant peptide MzDef may serve as a potential alternative antimicrobial and anticancer agent to be used in medicinal application.

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Characterization, cloning, expression and bioassay of vip3 gene isolated from an Egyptian Bacillus thuringiensis against whiteflies

Cited by 13 publications

References 22 publications

Vegetative Insecticidal Protein (Vip): A Potential Contender From Bacillus thuringiensis for Efficient Management of Various Detrimental Agricultural Pests

Vegetative Insecticidal Protein (Vip): A Potential Contender From Bacillus thuringiensis for Efficient Management of Various Detrimental Agricultural Pests

Expression of 1B capsid protein of Foot-and-mouth disease virus (FMDV) using baculovirus expression system and its validation in detecting SAT 2- specific antisera

Utilization of a recombinant defensin from Maize (Zea mays L.) as a potential antimicrobial peptide

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