PCR Analysis of cry7 Genes in Bacillus thuringiensis by the Five Conserved Blocks of Toxins

2001 ◽  
Vol 42 (2) ◽  
pp. 96-99 ◽  
Author(s):  
Eitan Ben-Dov ◽  
Robert Manasherob ◽  
Arieh Zaritsky ◽  
Ze'ev Barak ◽  
Yoel Margalith
Keyword(s):  
Bacillus Thuringiensis ◽  
Pcr Analysis

scholarly journals Characterization of Cry34/Cry35 Binary Insecticidal Proteins from Diverse Bacillus thuringiensis Strain Collections

2005 ◽  
Vol 71 (4) ◽  
pp. 1765-1774 ◽  
Author(s):  
H. Ernest Schnepf ◽  
Stacey Lee ◽  
JoAnna Dojillo ◽  
Paula Burmeister ◽  
Kristin Fencil ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Sequence Similarity ◽  
Diabrotica Virgifera Virgifera ◽  
Full Length ◽  
Pcr Analysis ◽  
Binding Motif ◽  
Diabrotica Virgifera ◽  
Crystal Proteins ◽  
Corn Rootworms ◽  
Geographic Origins

ABSTRACT Bacillus thuringiensis crystal proteins of the Cry34 and Cry35 classes function as binary toxins showing activity on the western corn rootworm, Diabrotica virgifera virgifera LeConte. We surveyed 6,499 B. thuringiensis isolates by hybridization for sequences related to cry35A genes, identifying 78 strains. Proteins of the appropriate molecular mass (ca. 44 kDa) for Cry35 were observed in 42 of the strains. Full-length, or nearly full-length, sequences of 34 cry34 genes and 16 cry35 genes were also obtained from cloning, PCR analysis, and DNA sequencing. These included representatives of all known Cry34A, Cry34B, Cry35A, and Cry35B classes, as well as a novel Cry34A/Cry35A-like pair. Bioassay analysis indicated that cry35-hybridizing strains not producing a ca. 14-kDa protein, indicative of Cry34, were not active on corn rootworms, and that the previously identified Cry34A/Cry35A pairs were more active than the Cry34B/Cry35B pairs. The cry35-hybridizing B. thuringiensis strains were found in locales and materials typical for other B. thuringiensis strains. Comparison of the sequences with the geographic origins of the strains showed that identical, or nearly identical, sequences were found in strains from both Australasia and the Americas. Sequence similarity searches revealed that Cry34 proteins are similar to predicted proteins in Photorhabdus luminescens and Dictyostelium discoidium, and that Cry35Ab1 contains a segment similar to beta-trefoil domains that may be a binding motif. The binary Cry34/Cry35 B. thuringiensis crystal proteins thus appear closely related to each other, are environmentally ubiquitous, and share sequence similarities consistent with activity through membrane disruption in target organisms.

scholarly journals Molecular typing of native Bacillus thuringiensis isolates from diverse habitats in India using REP-PCR and ERIC-PCR analysis

2012 ◽  
Vol 58 (2) ◽  
pp. 83-94 ◽  
Author(s):  
Jawahar Katara ◽  
Rupesh Deshmukh ◽  
Nagendra K. Singh ◽  
Sarvjeet Kaur
Keyword(s):  
Bacillus Thuringiensis ◽  
Molecular Typing ◽  
Pcr Analysis

scholarly journals Soybean plants expressing the Bacillus thuringiensis cry8-like gene show resistance to Holotrichia parallela

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Di Qin ◽  
Xiao-Yi Liu ◽  
Cristina Miceli ◽  
Qi Zhang ◽  
Pi-wu Wang
Keyword(s):  
Bacillus Thuringiensis ◽  
Transgenic Plants ◽  
Survival Rates ◽  
Pcr Analysis ◽  
Transgenic Soybean ◽  
Cry Protein ◽  
Transgenic Expression ◽  
Holotrichia Parallela ◽  
Soybean Plants ◽  
Soybean Leaves

Abstract Background Cry8-like from Bacillus thuringiensis (Bt) encodes an insecticidal crystal (Cry) protein. Holotrichia parallela (Coleoptera: Scarabaeoidae), commonly known as the dark black chafer, is a troublesome pest of soybean (Glycine max). To test whether cry8-like can confer resistance against H. parallela to soybean, we introduced cry8-like from the Bt strain HBF-18 into soybean cultivar Jinong 28. Results Quantitative reverse transcription-PCR analysis demonstrated that cry8-like was expressed most highly in soybean leaves. In addition, Southern blot assays revealed that one copy of the integrated fragment was present in the transformed plants. Eight independent cry8-like transgenic lines were subsequently fed on by H. parallela. Under H. parallela feeding stress, the survival rates of the non-transgenic plants were 92% lower than those of the transgenic plants. The mortality rate of H. parallela increased when the larvae fed on the roots of T1 transgenic soybean plants. Moreover, the surviving larvae were deformed, and their growth was inhibited. Conclusions Collectively, our data suggest that transgenic soybean plants expressing the cry8-like gene are more resistant to H. parallela than non-transgenic plants and that transgenic expression of the cry8-like gene may represent a promising strategy for engineering pest tolerance. The events generated in this study could thus be utilized in soybean breeding programs.

scholarly journals PCR analysis of the cryI insecticidal crystal family genes from Bacillus thuringiensis.

1994 ◽  
Vol 60 (1) ◽  
pp. 353-356 ◽  
Author(s):  
J Ceron ◽  
L Covarrubias ◽  
R Quintero ◽  
A Ortiz ◽  
M Ortiz ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Pcr Analysis

scholarly journals Fingerprinting of Bacillus thuringiensis Type Strains and Isolates by Using Bacillus cereus Group-Specific Repetitive Extragenic Palindromic Sequence-Based PCR Analysis

2005 ◽  
Vol 71 (3) ◽  
pp. 1346-1355 ◽  
Author(s):  
Arturo Reyes-Ramirez ◽  
Jorge E. Ibarra
Keyword(s):  
Bacillus Thuringiensis ◽  
Type Strain ◽  
Repeated Sequence ◽  
Antigenic Determinants ◽  
Pcr Analysis ◽  
Palindromic Sequence ◽  
Pcr Fingerprinting ◽  
Type Strains ◽  
Group Species ◽  
Repetitive Extragenic Palindromic

ABSTRACT A total of 119 Bacillus thuringiensis strains (83 type strains and 26 native isolates), as well as five B. cereus group species, were analyzed by repetitive extragenic palindromic sequence-based PCR analysis (Rep-PCR) fingerprinting. Primers Bc-REP-1 and Bc-REP-2 were specifically designed according to an extragenic 26-bp repeated sequence found in the six B. cereus group genomes reported. A total of 47 polymorphic bands were detected, and the patterns varied from 5 to 13 bands in number and from 0.2 to 3.8 kb in size. Virtually each type strain showed a distinctive B. cereus (Bc)-Rep-PCR pattern, except for B. thuringiensis serovars dakota (H serotype 15 [H15]) and sotto (H4a,4b), as well as serovars amagiensis (H29) and seoulensis (H35), which shared the same patterns. As expected, serovar entomocidus (H6) and its biovar subtoxicus showed an identical pattern; similarly, serovars sumiyoshiensis (H3a,3d) and fukuokaensis (H3a,3d,3e), which share two antigenic determinants, also showed identical Bc-Rep-PCR patterns. Interestingly, serovars israelensis (H14) and malaysiensis (H36), which share several phenotypic attributes, also showed identical Bc-Rep-PCR patterns. Native, coleopteran-active strains, including the self-agglutinated LBIT-74 strain, showed Bc-Rep-PCR patterns identical or very similar to that of the tenebrionis strain. Likewise, native mosquitocidal strains (including some self-agglutinated strains) also showed patterns identical or very similar to that of the serovar israelensis IPS-82 strain. Additionally, native β-exotoxin-producing strains from serovar thuringiensis showed patterns identical to that of the B. thuringiensis type strain. The B. cereus group-specific Bc-Rep-PCR fingerprinting technique was shown to be highly discriminative, fast, easy, and able to identify B. thuringiensis serotypes, including nonflagellar and self-agglutinated strains.

Isolation of Tn916-like conjugal elements from swine lot effluent

2000 ◽  
Vol 46 (6) ◽  
pp. 542-549 ◽  
Author(s):  
Bradley J Haack ◽  
Robert E Andrews Jr.
Keyword(s):  
Bacillus Thuringiensis ◽  
Enterococcus Faecalis ◽  
Tetracycline Resistance ◽  
Bacillus Species ◽  
Pcr Analysis ◽  
Soil Environment ◽  
Swine Waste ◽  
Chain Reaction ◽  
Plasmid Pc194 ◽  
Polymerase Chain

Isolates of Enterococcus faecalis obtained from a swine farrowing house outflow were examined for genetic elements similar to Tn916. Of the enterococci isolated, 71% were resistant to tetracycline. Among the tetracycline-resistant enterococci isolated from the outflow samples, approximately 34% were able to transfer the tetracycline resistance phenotype to Bacillus thuringiensis in cross-genus matings. The frequencies of transfer for 10 random isolates were comparable to those for transfer of Tn916 from E. faecalis to B. thuringiensis. In addition, these elements were shown to mobilize plasmid pC194 between Bacillus species, as did Tn916. Southern blot and polymerase chain reaction (PCR) analysis showed these elements share extensive structural homology with Tn916. The selected conjugal elements were capable of transfer to a Bacillus recipient in a soil environment. When the swine waste was introduced into the soil, the tetracycline resistant fecal enterococci levels rose from essentially undetectable levels to approximately 4 × 104and remained at this level for 4 weeks. After six months, including one winter, levels had decreased to 5 × 103.Key words: Enterococcus faecalis, Tn916, swine waste, genetic exchange, Bacillus thuringiensis, PCR.

scholarly journals Diversity of Bacillus thuringiensis Strains from Latin America with Insecticidal Activity against Different Mosquito Species

2003 ◽  
Vol 69 (9) ◽  
pp. 5269-5274 ◽  
Author(s):  
Jorge E. Ibarra ◽  
M. Cristina del Rincón ◽  
Sergio Ordúz ◽  
David Noriega ◽  
Graciela Benintende ◽  
...  
Keyword(s):  
Latin America ◽  
Bacillus Thuringiensis ◽  
Insecticidal Activity ◽  
Mosquito Species ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Pcr Analysis ◽  
Pcr Screening ◽  
Management Interventions ◽  
Plasmid Profiles

ABSTRACT The characterization of selected Bacillus thuringiensis strains isolated from different Latin America countries is presented. Characterization was based on their insecticidal activity against Aedes aegypti, Culex quinquefasciatus, and Anopheles albimanus larvae, scanning electron microscopy, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and plasmid profiles as well as PCR analysis using novel general and specific primers for cry and cyt genes encoding proteins active against mosquitoes (cyt1, cyt2, cry2, cry4A, cry4B, cry10, cry11, cry17, cry19, cry24, cry25, cry27, cry29, cry30, cry32, cry39, and cry40). Strains LBIT315, LBIT348, and IB604 showed threefold higher mosquitocidal activity against A. aegypti and C. quinquefasciatus larvae than B. thuringiensis subsp. israelensis and displayed high similarities with the B. thuringiensis subsp. israelensis used in this study with regard to protein and plasmid profiles and the presence of cry genes. Strain 147-8906 has activity against A. aegypti similar to that of B. thuringiensis subsp. israelensis but has different protein and plasmid profiles. This strain, harboring cry11, cry30, cyt1, and cyt2 genes, could be relevant for future resistance management interventions. Finally, the PCR screening strategy presented here led us to identify a putative novel cry11B gene.

Distribution, Serological Identification, and PCR Analysis of Bacillus thuringiensis Isolated from Soils of Korea

1998 ◽  
Vol 37 (3) ◽  
pp. 195-200 ◽  
Author(s):  
Ho San Kim ◽  
Dae Won Lee ◽  
Soo Dong Woo ◽  
Yong Man Yu ◽  
Seok Kwon Kang
Keyword(s):  
Bacillus Thuringiensis ◽  
Pcr Analysis ◽  
Serological Identification

Detection of H. Pylori by PCR analysis in liver biopsy specimens

2001 ◽  
Vol 34 (0) ◽  
pp. 185-186
Author(s):  
I Kupcinskas
Keyword(s):  
Liver Biopsy ◽  
Pcr Analysis ◽  
Biopsy Specimens ◽  
H Pylori

Characterization of Partial Gene Deletions in Type III von Willebrand Disease with Alloantibody Inhibitors

1994 ◽  
Vol 72 (02) ◽  
pp. 180-185 ◽  
Author(s):  
David J Mancuso ◽  
Elodee A Tuley ◽  
Ricardo Castillo ◽  
Norma de Bosch ◽  
Pler M Mannucci ◽  
...  
Keyword(s):  
Von Willebrand Factor ◽  
Von Willebrand Disease ◽  
Pcr Analysis ◽  
Gene Deletions ◽  
Factor Gene ◽  
Type Iii ◽  
Large Deletions ◽  
Von Willebrand ◽  
Willebrand Factor

Summaryvon Willebrand factor gene deletions were characterized in four patients with severe type III von Willebrand disease and alloantibodies to von Willebrand factor. A PCR-based strategy was used to characterize the boundaries of the deletions. Identical 30 kb von Willebrand factor gene deletions which include exons 33 through 38 were identified in two siblings of one family by this method. A small 5 base pair insertion (CCTGG) was sequenced at the deletion breakpoint. PCR analysis was used to detect the deletion in three generations of the family, including two family members who are heterozygous for the deletion. In a second family, two type III vWD patients, who are distant cousins, share an -56 kb deletion of exons 22 through 43. The identification and characterization of large vWF gene deletions in these type III vWD patients provides further support for the association between large deletions in both von Willebrand factor alleles and the development of inhibitory alloantibodies.

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