Isolation, Cloning, DNA Sequencing and Bioinformatics Analysis of the Parasporin–1 Gene of Bacillus thuringiensis

Assaeedi, Abdulrahman; Osman, Gamal

doi:10.4172/jpb.1000435

Cited by 3 publications

(3 citation statements)

References 23 publications

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“…The Cry1Ac-binding APN proteins have also been purified from the BBMVs of Lymantria dispar , Heliothis virescens, and Plutella xylostella (Gill et al, 1995, Palma et al, 2014). However, few reports suggest that not all APN surface enzymes act as toxin binding proteins (Maaty, 1999, Assaeedi and Osman, 2017). From the current and earlier findings, it may be inferred that the interference with the midgut function seems to be the common mechanism by which most effective insecticidal proteins, including the Vip3Aa toxin, kill their target insect.…”

Section: Discussionmentioning

confidence: 99%

First report of detection of the putative receptor of Bacillus thuringiensis toxin Vip3Aa from black cutworm (Agrotis ipsilon)

Osman

Altaf

Saleh

et al. 2018

Saudi Journal of Biological Sciences

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Black cutworm (BCW) , an economically important lepidopteran insect, has attracted a great attention. (Bt) is spore forming soil bacteria and is an excellent environment-friendly approach for the control of phytophagous and disease-transmitting insects. In fact, bio-pesticide formulations and insect resistant transgenic plants based on the bacterium Bt delta-endotoxin have attracted worldwide attention as a safer alternative to harmful chemical pesticides. The major objective of the current study was to understand the mechanism of interaction of Bt toxin with its receptor molecule(s). The investigation involved the isolation, identification, and characterization of a putative receptor - vip3Aa. In addition, the kinetics of vip toxin binding to its receptor molecule was also studied. The present data suggest that Vip3Aa toxin bound specifically with high affinity to a 48-kDa protein present at the brush border membrane vesicles (BBMV) prepared from the midgut epithelial cells of BCW larvae.

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Section: Discussionmentioning

confidence: 99%

First report of detection of the putative receptor of Bacillus thuringiensis toxin Vip3Aa from black cutworm (Agrotis ipsilon)

Osman

Altaf

Saleh

et al. 2018

Saudi Journal of Biological Sciences

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“…In a study, the parasporin 1 gene of Bacillus thuringiensis was amplified by PCR and cloned into pGEM-vector. The predicted protein encoded by the 2371 nucleotides’ long gene sequence was composed of 789 amino acids with an estimated molecular weight of 84 kDa [ 15 ]. Vector pRham-SUMO was used to clone and sequence the nagH gene of C. chauvoei [ 16 ].…”

Section: Discussionmentioning

confidence: 99%

Cloning and sequence analysis of a partial CDS of leptospiral ligA gene in pET-32a - Escherichia coli DH5α system

Soman¹,

Mini²,

Joseph³

et al. 2018

Vet World

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AimThis study aims at cloning, sequencing, and phylogenetic analysis of a partial CDS of ligA gene in pET-32a - Escherichia coli DH5α system, with the objective of identifying the conserved nature of the ligA gene in the genus Leptospira.Materials and MethodsA partial CDS (nucleotide 1873 to nucleotide 3363) of the ligA gene was amplified from genomic DNA of Leptospira interrogans serovar Canicola by polymerase chain reaction (PCR). The PCR-amplified DNA was cloned into pET-32a vector and transformed into competent E. coli DH5α bacterial cells. The partial ligA gene insert was sequenced and the nucleotide sequences obtained were aligned with the published ligA gene sequences of other Leptospira serovars, using nucleotide BLAST, NCBI. Phylogenetic analysis of the gene sequence was done by maximum likelihood method using Mega 6.06 software.ResultsThe PCR could amplify the 1491 nucleotide sequence spanning from nucleotide 1873 to nucleotide 3363 of the ligA gene and the partial ligA gene could be successfully cloned in E. coli DH5α cells. The nucleotide sequence when analyzed for homology with the reported gene sequences of other Leptospira serovars was found to have 100% homology to the 1910 bp to 3320 bp sequence of ligA gene of L. interrogans strain Kito serogroup Canicola. The predicted protein consisted of 470 aminoacids. Phylogenetic analysis revealed that the ligA gene was conserved in L.interrogans species.ConclusionThe partial ligA gene could be successfully cloned and sequenced from E. coli DH5α cells. The sequence showed 100% homology to the published ligA gene sequences. The phylogenetic analysis revealed the conserved nature of the ligA gene. Further studies on the expression and immunogenicity of the partial LigA protein need to be carried out to determine its competence as a subunit vaccine candidate.

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“…Furthermore, Cry15Aa is highly homologous to mosquitocidal Mtx2 and Mtx3 proteins from B. sphaericus [9,10]. Interestingly, these mosquitocidal proteins show structural similarities with β-barrel-lined membrane pore-forming toxin, aerolysin from Aeromonas hydrophila [11][12][13]. The 37 kDa precursor cleaves into a 30 kDa active form through proteolytic processing at both N-and C-terminus and the activated toxin exhibits high cytotoxicity to cancer cells.…”

Section: Parasporins (Ps) Have Been Identified As Cry Bacterial Cytocmentioning

confidence: 99%

Overexpression and Cytocidal Activity of Parasporin-1 from Bacillus thuringiensis against Human Cancer Cell Lines

Assaeedi¹,

Osman²

2017

J Microb Biochem Technol

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Isolation, Cloning, DNA Sequencing and Bioinformatics Analysis of the Parasporin–1 Gene of Bacillus thuringiensis

Cited by 3 publications

References 23 publications

First report of detection of the putative receptor of Bacillus thuringiensis toxin Vip3Aa from black cutworm (Agrotis ipsilon)

First report of detection of the putative receptor of Bacillus thuringiensis toxin Vip3Aa from black cutworm (Agrotis ipsilon)

Cloning and sequence analysis of a partial CDS of leptospiral ligA gene in pET-32a - Escherichia coli DH5α system

Overexpression and Cytocidal Activity of Parasporin-1 from Bacillus thuringiensis against Human Cancer Cell Lines

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