Improved shuttle vector for expression of chitinase gene in Bacillus thuringiensis

Lertcanawanichakul, M.; Wiwat, Chanpen

doi:10.1046/j.1365-2672.2000.00777.x

Cited by 14 publications

(9 citation statements)

References 17 publications

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“…When the recombinant bacterium was cultivated in NB without erythromycin with daily transfers for 31 days, 90% of erythromycin resistant colonies were obtained. The high segregational stability of pEBchiA74 suggests that this recombinant could be stably maintained after several passages, similar to the pBCX shuttle vector (Lertcanawanichakul and Wiwat 2000), or pHYA2 and pEHchiA74 harboring chitinase genes (Lertcanawanichakul et al 2004;CasiqueArroyo et al 2007). …”

Section: Resultsmentioning

confidence: 97%

Hyperproduction of chitinase influences crystal toxin synthesis and sporulation of Bacillus thuringiensis

Barboza‐Corona

Ortiz-Rodríguez

Fuente-Salcido

et al. 2009

Antonie van Leeuwenhoek

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Bacillus thuringiensis HD-73 was transformed with the endochitinase gene chiA74 under the control of a strong promoter (pcytA) and a 5' mRNA stabilizing (STAB-SD) sequence (HD-73-pEBchiA74). Expression levels were compared with those observed from the wild type strain (HD-73) and the recombinant HD-73 strain expressing chiA74 under the control of its native promoter (HD-73-pEHchiA74). The chitinolytic activity of HD-73-pEBchiA74 was markedly elevated, being ~58- and 362-fold higher than, respectively, HD-73-pEHchiA74 and parental HD-73, representing the highest levels of chitinase expression in recombinant B. thuringiensis reported to date. Parasporal crystals measured under transmission electron microscopy showed that HD-73 produced crystals of 1.235 (+/-0.214) and 1.356 (+/-0.247) mum in length when the bacterium was grown in respectively, NBS and NBS with glucose. Otherwise, HD-73-pEBchiA74 synthesized crystals of 1.250 (+/-0.222) and 1.139 (+/-0.202) mum in length when cultivated in NBS and NBS with glucose, respectively, values that showed a diminution of ~10 and 20% compared with crystals produced by HD-73-pEHchiA74 grown under the same conditions. Comparison of viable spore counts per ml showed that HD-73-pEBchiA74 produced fewest viable spores (1.5 x 10(9), 1.3 x 10(9)), compared to HD-73-pEHchiA74 (4.9 x 10(9), 5.3 x 10(9)) and HD-73 (6.8 x 10(9), 8.8 x 10(9)) when grown in NBS and NBS supplemented with glucose, respectively. No change in cellular protease activity was observed despite the overproduction of the chitinase.

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Section: Resultsmentioning

confidence: 97%

Hyperproduction of chitinase influences crystal toxin synthesis and sporulation of Bacillus thuringiensis

Barboza‐Corona

Ortiz-Rodríguez

Fuente-Salcido

et al. 2009

Antonie van Leeuwenhoek

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“…It is evident from the literature that chitinases play a synergistic role in enhancing entomotoxicity of biocontrol agents [37,38]. Brar et al [38] demonstrated that chitinases play synergistic role in Bacillus thuringiensis ssp.…”

Section: Validation Of Optimal Conditions and Optimal Responses In Somentioning

confidence: 98%

Bioproduction of hydrolytic enzymes using apple pomace waste by A. niger: applications in biocontrol formulations and hydrolysis of chitin/chitosan

Dhillon

Brar

Valero

et al. 2011

Bioprocess Biosyst Eng

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Studies were carried out for β-glucosidase production using apple pomace (AP) in solid-state fermentation using 2(4) factorial design and response surface methodology. The influence of four independent variables including initial moisture level and inducers [veratryl alcohol (VA), lactose (LAC) and copper sulfate (CS)] was studied. The experimental design showed that initial moisture level had significant negative effect on the response. Higher β-glucosidase activity of 64.18 IU/gram fermented substrate (gfs) was achieved in solid-state tray fermentation with optimum conditions having initial moisture level 55% (v/w), pH 4.5, 2 mM/kg VA, 2% (w/w) LAC and 1.5 mM/kg CS concentration, respectively,. The non-specific chitinase 70.28 ± 6.34 IU/gfs and chitosanase activities 60.18 ± 6.82 to 64.20 ± 7.12 IU/gfs were observed. The study demonstrated that AP can be potentially used for the β-glucosidase production by Aspergillus niger. Moreover, β-glucosidase can be used for the hydrolysis of chitin/chitosan to depolymerized products and in the formulation of biocontrol agents for enhanced entomotoxicity levels.

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“…To tackle the difficulties of stable expression of the chitinase gene in B. thuringiensis, the shuttle vector pBcX was constructed by recombining the 2.56-kb fragment of plasmid pBluescript II KS with plasmid pBc16. This is useful in cloning foreign DNA fragments (Lertcanawanichakul and Wiwat 2000). Sun et al used a shuttle vector constructed using the small plasmid pBMB2062 cloned from B. thuringiensis subsp kurstaki strain YBT-1520 and was found capable of stabilizing the inheritance and expression of insecticidal crystal proteins (Sun et al 2000).…”

Section: Shuttle Plasmid Vectors In the B Cereus Groupmentioning

confidence: 98%

Construction and Application in Plasmid Vectors of Bacillus cereus Group

Wang

et al. 2012

Bacillus Thuringiensis Biotechnology

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The B. cereus group ( B. cereus sense lato) is a subgroup of relatedBacillus containing six species: B. thuringiensis, B. anthracis, B. cereus, B. weihenstephanensis, B. mycoides and B. pseudomycoides. The first three species are opportunistic or pathogenic to insects or mammals, whereas the last three are generally regarded as nonpathogenic. Different types of plasmid vectors play an important role in studying gene function in genetically engineered bacteria or construction of mutant library. The researchers have constructed a variety of shuttle vectors in two or more different host strains. Expression vectors are widely used in construction of highly effective engineered strains. Resolution vectors can site-specifically transfer recombinant genes. Integration vectors could integrate foreign genes into the chromosome or plasmid of host strains and are free of antibiotic resistance genes, therefore eliminating security risks. In recent years, researchers have made great progress in the development of genetic manipulation vectors. Here we summarize latest progress in vector construction and their application in Bacillus cereus group.

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Improved shuttle vector for expression of chitinase gene in Bacillus thuringiensis

Cited by 14 publications

References 17 publications

Hyperproduction of chitinase influences crystal toxin synthesis and sporulation of Bacillus thuringiensis

Hyperproduction of chitinase influences crystal toxin synthesis and sporulation of Bacillus thuringiensis

Bioproduction of hydrolytic enzymes using apple pomace waste by A. niger: applications in biocontrol formulations and hydrolysis of chitin/chitosan

Construction and Application in Plasmid Vectors of Bacillus cereus Group

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