RAP-PCR fingerprinting reveals time-dependent expression of development-related genes following differentiation process of Bacillus thuringiensis

Gene expression profiles are important data to reveal gene functions putatively involved in crucial biological processes. RNA arbitrarily primed-polymerase chain reaction (RAP-PCR) and specifically primed reverse transcription PCR (RT-PCR) were combined to screen differentially expressed genes following development of a commercial Bacillus thuringiensis (Bt) subsp. kurstaki strain 8010 (serotype 3a3b). Six differentially expressed transcripts (RAP1 to RAP6) were obtained. The RAP1 encoded a putative triple helix repeat-containing collagen or an exosporium protein H related to spore pathogenicity. The RAP2 was homologous to a ClpX protease and an ATP-dependent protease La (LonB) likely acted as virulence factors. The RAP3 was homologous to a beta subunit of propionyl-CoA carboxylase (PCC) enzyme (PCCB) required for the development of Myxococcus xanthus. The RAP4 had homology to a quinone oxidoreductase involved in electron transport and ATP formation. The RAP5 showed significant homology to a uridine kinase (UDK) mediated phosphorylation of uridine, azauridine. The RAP6 shared high sequence identity with 3-methyl-2-oxobutanoate-hydroxymethyltransferase (MOHMT) (EC 2.1.2.11) [also known as ketopantoate hydroxymethyltransferase (KPHMT) or PanB] involved in the operation of the tricarboxylic acid cycle. The findings described here would help to elucidate the molecular mechanisms underlying the differentiation process of Bt and unravel novel pathogenic genes.The accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author

Characterization of cry1Cb3 and cry1Fb7 from Bacillus thuringiensis subsp. galleriae

Two cry1-type genes encoding insecticidal crystal proteins (ICPs) were detected by PCR-RFLP and cloned from Bacillus thuringiensis subsp. galleriae 87. The nucleotide sequences were deposited in GenBank with accession numbers EU679501 and EU679502, and designated as cry1Fb7 and cry1Cb3 respectively by B. thuringiensis Delta- Endotoxin Nomenclature Committee. cry1Cb3 shared 99% homology with other cry1Cb genes. The existence of two additional stop codons indicated cry1Cb3 was a silent gene. The cry1Cb3 was 3531 bp with 38.98% G+C content and its first open reading frame (ORF) encoded a protein of 213 amino acid residues with a calculated molecular weight of 23.8 kDa and a predicted pI value of 4.63. Five amino acid sequence blocks (block 1, block 2, block 3, block 4 and block 5) were found in Cry1Cb3. Translation of cry1Fb7 revealed an ORF of 3525 bp with 39.12% G+C content and a protein with a calculated molecular weight of 133.2 kDa and a predicted pI value of 5.18. Cry1Fb7 had five amino acid sequence blocks (blocks 1, 2, 3, 4 and 5) and three domains (I, II and III), which consisted of 218 residues (Leu34 to Ala252), 197 residues (Thr257 to Asp454), and 138 residues (Ile464 to Glu600), respectively

Towards Sustainable Green Adjuvants for Microbial Pesticides: Recent Progress, Upcoming Challenges, and Future Perspectives

Microbial pesticides can be significantly improved by adjuvants. At present, microbial pesticide formulations are mainly wettable powders and suspension concentrations, which are usually produced with adjuvants such as surfactants, carriers, protective agents, and nutritional adjuvants. Surfactants can improve the tension between liquid pesticides and crop surfaces, resulting in stronger permeability and wettability of the formulations. Carriers are inert components of loaded or diluted pesticides, which can control the release of active components at appropriate times. Protective agents are able to help microorganisms to resist in adverse environments. Nutritional adjuvants are used to provide nutrients for microorganisms in microbial pesticides. Most of the adjuvants used in microbial pesticides still refer to those of chemical pesticides. However, some adjuvants may have harmful effects on non-target organisms and ecological environments. Herein, in order to promote research and improvement of microbial pesticides, the types of microbial pesticide formulations were briefly reviewed, and research progress of adjuvants and their applications in microbial pesticides were highlighted, the challenges and the future perspectives towards sustainable green adjuvants of microbial pesticides were also discussed in this review

Towards Sustainable Green Adjuvants for Microbial Pesticides: Recent Progress, Upcoming Challenges, and Future Perspectives

Microbial pesticides can be significantly improved by adjuvants. At present, microbial pesticide formulations are mainly wettable powders and suspension concentrations, which are usually produced with adjuvants such as surfactants, carriers, protective agents, and nutritional adjuvants. Surfactants can improve the tension between liquid pesticides and crop surfaces, resulting in stronger permeability and wettability of the formulations. Carriers are inert components of loaded or diluted pesticides, which can control the release of active components at appropriate times. Protective agents are able to help microorganisms to resist in adverse environments. Nutritional adjuvants are used to provide nutrients for microorganisms in microbial pesticides. Most of the adjuvants used in microbial pesticides still refer to those of chemical pesticides. However, some adjuvants may have harmful effects on non-target organisms and ecological environments. Herein, in order to promote research and improvement of microbial pesticides, the types of microbial pesticide formulations were briefly reviewed, and research progress of adjuvants and their applications in microbial pesticides were highlighted, the challenges and the future perspectives towards sustainable green adjuvants of microbial pesticides were also discussed in this review

From Detection to Resection: Photoacoustic Tomography and Surgery Guidance with Indocyanine Green Loaded Gold Nanorod@liposome Core–Shell Nanoparticles in Liver Cancer

Conventional imaging methods encounter challenges in diagnosing liver cancer that is less than 10 mm or without typical hypervascular features. With deep penetration and high spatial resolution imaging capability, the emerging photoacoustic tomography may offer better diagnostic efficacy for noninvasive liver cancer detection. Moreover, near-infrared fluorescence imaging-guided hepatectomy was proven to be able to identify nodules at the millimeter level. Thus, suitable photoacoustic and fluorescence dual-modality imaging probe may benefit patients in early diagnosis and complete resection. In this study, we fabricated indocyanine green loaded gold nanorod@liposome core–shell nanoparticles (Au@liposome–ICG) to integrate both imaging strategies. These nanoparticles exhibit superior biocompatibility, high stability, and enhanced dual-model imaging signals. Next, we explored their effectiveness of tumor detection and surgery guidance in orthotopic liver cancer mouse models. Histological analysis confirmed the accuracy of the probe in liver cancer detection and resection. This novel dual-modality nanoprobe holds promise for early diagnosis and better surgical outcome of liver cancer and has great potential for clinical translation

Cry11Aa Interacts with the ATP-Binding Protein from Culex quinquefasciatus To Improve the Toxicity

Cry11Aa displays high toxicity to the larvae of several mosquito species, including Aedes, Culex, and Anopheles. To study its binding characterization against Culex quinquefasciatus, Cry11Aa was purified and western blot results showed that Cry11Aa could bind successfully to the brush border membrane vesicles. To identify Cry11Aa-binding proteins in C. quinquefasciatus, a biotin-based protein pull-down experiment was performed and seven Cry11Aa-binding proteins were isolated from the midgut of C. quinquefasciatus larvae. Analysis of liquid chromatography–tandem mass spectrometry showed that one of the Cry11Aa-binding proteins is the ATP-binding domain 1 family member B. To investigate its binding property and effect on the toxicity of Cry11Aa, western blot, far-western blot, enzyme-linked immunosorbent assay, and bioassays of Cry11Aa in the presence and absence of the recombinant ATP-binding protein were performed. Our results showed that the ATP-binding protein interacted with Cry11Aa and increased the toxicity of Cry11Aa against C. quinquefasciatus. Our study suggests that midgut proteins other than the toxin receptors may modulate the toxicity of Cry toxins against mosquitoes