Characterization ofBacillus thuringiensismutants and natural isolates by molecular methods

1997 ◽  
Vol 43 (5) ◽  
pp. 403-410 ◽  
Author(s):  
Yong Chul Jung ◽  
Sung Uk Kim ◽  
Song Hae Bok ◽  
Ho Yong Park ◽  
Jean-Charles Côté ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Bacillus Thuringiensis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Rice Grain ◽  
Chain Reaction ◽  
Hyphantria Cunea ◽  
Natural Isolates ◽  
Grain Dust ◽  
Polymerase Chain

Two Bacillus thuringiensis var. kurstaki HD-1 mutants, two Bacillus thuringiensis var. israelensis HD-500 mutants, and four rice grain dust isolates were characterized using microscopic examination and protein profiles of purified crystals on sodium dodecyl sulfate – polyacrylamide gel electrophoresis. Specific detection of cryI- and cryIV-type genes was performed in a polymerase chain reaction using cryI and cryIV-specific oligonucleotide primers. The cry-type genes under study consisted of cryIA(a), cryI(A)b, cryI(A)c, cryIB, and cryIV. Presence or absence of the cryI- and cryIV-type genes was further confirmed by Southern blotting followed by hybridization with specific cryI and cryIV gene fragments. A genetically modified strain of B. thuringiensis var. kurstaki HD-1, called OZK-13 and obtained following mutagenesis with ozone, was shown to contain cryIA(a), cryIA(b), and cryIA(c) genes. Another kurstaki HD-1 mutant, called NGK-13 and obtained following treatment with N-methyl-N′-nitro-N-nitrosoguanidine (MNNG), was shown to have lost the cryIA(b) gene while retaining the cryIA(a) and cryIA(c) genes. NGI-23-1, an oligosporogenous–multicrystalliferous mutant of B. thuringiensis var. israelensis (Bti) HD-500, obtained following treatment with MNNG contained cryIV-type genes. NGI-22, an oligosporogenous–acrystalliferous mutant of Bti HD-500, contained no cryI- nor cryIV-type genes. The rice grain dust isolate BT-285 contained the cryIA(a) and cryIA(c) genes. Isolate BT-14 contained only the cryIA(c) gene, whereas isolate BT-209 contained cryIA(a), cryIA(b), and cryIB genes. Isolate BT-205 contained no cryI- nor cryIV-type genes. Bacillus thuringiensis mutants and natural isolates shown to contain cryI-type genes were tested for their insecticidal activities in a series of bioassays against Hyphantria cunea Drury (Lepidoptera: Arctiidae). All cryI-carrying strains were toxic against the insect larvae. BT-205 was also tested and exhibited no toxicity against the insect larvae.Key words: Bacillus thuringiensis, δ-endotoxin crystal, cry-type genes, polymerase chain reaction.

scholarly journals ANALYSIS OF THE INHIBITORY POTENTIAL OF STREPTOCOCCUS SALIVARIUS ISOLATED FROM ADULT SALIVA AND THE TONGUE DORSUM FOR THE GROWTH OF CANDIDA ALBICANS

2017 ◽  
Vol 9 ◽  
pp. 3
Author(s):  
Nindy Fairiska ◽  
Boy M Bachtiar ◽  
Ferry P Gultom
Keyword(s):  
Polymerase Chain Reaction ◽  
Candida Albicans ◽  
Sodium Dodecyl Sulfate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Streptococcus Salivarius ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Inhibitory Potential

Objectives: The objective of this study is to analyze the effectiveness of Streptococcus salivarius and its protein for inhibiting the growth of Candidaalbicans.Methods: The analysis was conducted using polymerase chain reaction, sodium dodecyl sulfate polyacrylamide gel electrophoresis, a Bradford test,deferred antagonism test, and well-diffusion agar.Result: S. salivarius, isolated from saliva and the tongue dorsum, and its protein do not inhibit the growth of C. albicans. The morphology of C. albicansdid not change after being exposed to protein produced by S. salivarius.Conclusions: S. salivarius and its protein do not inhibit the growth of C. albicans. However, the bacterium has the capacity to maintain fungusmorphology in the form of blastospora.

scholarly journals Evaluation of a Polymerase Chain Reaction-Based Heteroduplex Assay for Detecting the Adulteration of Processed Orange Juice with Mandarin Juice

2006 ◽  
Vol 89 (4) ◽  
pp. 1052-1060 ◽  
Author(s):  
Rachel Mooney ◽  
Louise Chappell ◽  
Angus I Knight
Keyword(s):  
Polymerase Chain Reaction ◽  
Orange Juice ◽  
Polyacrylamide Gel Electrophoresis ◽  
Pcr Amplification ◽  
Intergenic Spacer ◽  
Target Sequence ◽  
Chain Reaction ◽  
Good Repeatability ◽  
Polymerase Chain ◽  
Mandarin Juice

Abstract A polymerase chain reaction (PCR)-based heteroduplex assay was evaluated for the detection of mandarin juice in processed orange juice. PCR amplification of a fragment of the chloroplast trnT-trnL intergenic spacer derived from mixtures of DNA extracted from orange and mandarin juice resulted in heteroduplex formation. The heteroduplex resulted from the co-amplification of a fragment containing an 8 base-pair indel that distinguished mixtures of orange and mandarin juice from orange juice and mandarin juice alone. The heteroduplex assay was evaluated against authentic juices obtained from different citrus species and confirmed that the marker was homogeneous within Citrus. The data obtained demonstrated maternal inheritance of chloroplast type in Citrus sp. and allowed the identification and confirmation of the maternal parentage of unknown and known citrus hybrids. Analysis of the quantitative potential of the PCR and polyacrylamide gel electrophoresis (PAGE) analysis demonstrated good repeatability with a coefficient of variation of 7.5%. Greatest sources of variance in experimental results were attributable to species and varietal differences in the levels of the PCR target. Mandarin juice contained approximately 18% (w/v) less PCR target sequence than did orange juice. The assay was tested in a blind trial using processed juices and correctly identified 20/22 samples with no false-positive results.

scholarly journals Application of Polymerase Chain Reaction–Single Strand Conformational Polymorphism (PCR–SSCP) to Identification of Flatfish Species

1999 ◽  
Vol 82 (4) ◽  
pp. 903-907 ◽  
Author(s):  
Ana Céspedes ◽  
Teresa García ◽  
Esther Carrera ◽  
Isabel González ◽  
Esther Carrera ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Polyacrylamide Gel Electrophoresis ◽  
Single Strand ◽  
Chain Reaction ◽  
Greenland Halibut ◽  
Single Strand Conformational Polymorphism ◽  
Conformational Polymorphism ◽  
Flatfish Species ◽  
Mitochondrial Cytochrome B ◽  
Polymerase Chain

Abstract A method of DNA analysis based on polymerase chain reaction–single strand conformational polymorphism (PCR–SSCP) was developed to verify the authenticity of labeled raw and frozen fillets of some flatfish species. PCR was used to amplify a short fragment (201 bp) of the mitochondrial cytochrome b gene, which was denatured and analyzed by native polyacrylamide gel electrophoresis for detection of SSCPs. Species-specific patterns of DNA bands were obtained for sole (Solea solea), European plaice (Pleuronectes platessa), flounder (Platichthys flesus), and Greenland halibut (Reinhardtius hippoglossoides).

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