scholarly journals Antibacterial Mechanism of Gloverin2 from Silkworm, Bombyx mori

2018 ◽  
Vol 19 (8) ◽  
pp. 2275 ◽  
Author(s):  
Qian Wang ◽  
Pengchao Guo ◽  
Zhan Wang ◽  
Huawei Liu ◽  
Yunshi Zhang ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Expression System ◽  
Random Coil ◽  
Antibacterial Mechanism ◽  
Cell Integrity ◽  
Aggregation State ◽  
Coil Structure ◽  
Prokaryotic Expression System ◽  
Escherichia Coli Jm109 ◽  
Α Helix

Gloverin is one of the glycine-rich antimicrobial peptide exclusively found in Lepidoptera insects. It is generally activated through the innate immune system in insects. In this study, recombinant Gloverin2 from Bombyx mori (BmGlv2) was synthesized using a prokaryotic expression system. Circular dichroism spectroscopy showed that the recombinant BmGlv2 has random coil structure, which is relatively stable at the temperatures ranging from 15 to 82.5 °C. Antimicrobial activity analysis revealed that BmGlv2 significantly inhibited the growth of gram-negative bacteria, Escherichia coli JM109 and Pseudomonas putida, by disrupting cell integrity. Western blotting and immunofluorescence analyses suggested that BmGlv2 absorbed on the cell surface after incubation, which might be the first step in the antibacterial process. Our results also proved that the cell wall component lipopolysaccharides (LPS) induce a conformational change in BmGlv2 from a random coil to α-helix. Subsequently, α-helical BmGlv2 would recruit more BmGlv2 and form higher aggregation state. Collectively, these findings expand our understanding of antibacterial mechanism of BmGlv2.

A Comparison Study of the Effects of Ultrasonication on the Gelation Behavior between the Silk Fibroin of Domestic and Wild Silkworms

2014 ◽  
Vol 941-944 ◽  
pp. 989-993
Author(s):  
Yuan Zhou
Keyword(s):  
Bombyx Mori ◽  
Structural Transformation ◽  
Gelation Time ◽  
Random Coil ◽  
Ultrasonic Power ◽  
Comparison Study ◽  
Silk Fibroins ◽  
Gelation Behavior ◽  
Α Helix ◽  
Β Sheet

This paper was concerned with the effects of ultrasonication on the gelation behavior of silk fibroins (SF), and a comparison of Domestic and Wild silkworms was studied. The results show that: with the increase of ultrasonic power, the gelation time of domestic (Bombyx Mori) SF solution decreased sharply. But wild silkworms (Antheraea yamamai and Antheraea pernyi) SF were different, When the power of utrasonication was lower than 400-500 W, the velocity of gelation were accelerated, and when the ultrasonic power was higher than 400-500W, the gelation time were delayed. Whatever domestic or wild silkworms, the mechanism of the effects of ultrasonication on the gelation behavior was that the ultrasonication promoted the structural transformation of SF molecules from random coil or α-helix to β-sheet.

Characterization of Putative a (1,3)-β-D-glucan (curdlan) Synthase for a Low Molecular Weight Curdlan Biosynthesis from Agrobacterium sp. M503

2013 ◽  
Vol 807-809 ◽  
pp. 2031-2034
Author(s):  
Yu Mei Li ◽  
Qiang Li ◽  
Sheng Han ◽  
Dong Xue Song ◽  
Yan Hong Qu ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Random Coil ◽  
Low Molecular Weight ◽  
Base Pairs ◽  
Reading Frame ◽  
Coil Structure ◽  
Α Helix ◽  
Encoding Protein ◽  
Β Sheet

A β-(1,3)-D-glucan (curdlan) synthase gene for a low molecular weight curdlan biosynthesis, crdSAg, from Agrobacterium sp. M503 was cloned and its encoding protein was characterized by several online protein analysis softwares. The crdSAg consists of 1965-base-pairs Open Reading Frame (ORF) encoding a protein with molecular weight approximate 73.5 kDa, which contains the conserved domain of CESA-CelA_like belonging to glycosyltransferase family 2 (GT2). Moreover, CrdSAg was a membrane protein with seven hydrophobic transmembrance domains. The second structure analysis indicated it was composed of 43.12% α-helix, 17.89% β-sheet, and 38.99% random coil structure. These data will lay a foundation to clarify the biosynthesis mechanism of the low molecular weight curdlan.

scholarly journals Bombyx mori Pupae Efficiently Produce Recombinant AAV2/HBoV1 Vectors with a Bombyx mori Nuclear Polyhedrosis Virus Expression System

Viruses ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 704
Author(s):  
Qian Yu ◽  
Pengfei Chang ◽  
Xiaoxuan Liu ◽  
Peng Lü ◽  
Qi Tang ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Bombyx Mori ◽  
Nuclear Polyhedrosis Virus ◽  
Expression System ◽  
Efficient Production ◽  
Human Bocavirus ◽  
Adeno Associated Virus ◽  
Silkworm Pupae ◽  
Polyhedrosis Virus ◽  
Nuclear Polyhedrosis

Recombinant adeno-associated virus (AAV) vectors have broad application prospects in the field of gene therapy. The establishment of low-cost and large-scale manufacturing is now the general agenda for industry. The baculovirus-insect cell/larva expression system has great potential for these applications due to its scalability and predictable biosafety. To establish a more efficient production system, Bombyx mori pupae were used as a new platform and infected with recombinant Bombyx mori nuclear polyhedrosis virus (BmNPV). The production of a chimeric recombinant adeno-associated virus (rAAV) serotype 2/human bocavirus type-1 (HBoV1) vector was used to evaluate the efficiency of this new baculovirus expression vector (BEV)–insect expression system. For this purpose, we constructed two recombinant BmNPVs, which were named rBmNPV/AAV2Rep-HBoV1Cap and rBmNPV/AAV2ITR-eGFP. The yields of rAAV2/HBoV1 derived from the rBmNPV/AAV2Rep-HBoV1Cap and rBmNPV/AAV2ITR-eGFP co-infected BmN cells exceeded 2 × 104 vector genomes (VG) per cell. The rBmNPV/AAV2Rep-HBoV1Cap and rBmNPV/AAV2ITR-eGFP can express stably for at least five passages. Significantly, rAAV2/HBoV1 could be efficiently generated from BmNPV-infected silkworm larvae and pupae at average yields of 2.52 × 1012 VG/larva and 4.6 × 1012 VG/pupa, respectively. However, the vectors produced from the larvae and pupae had a high percentage of empty particles, which suggests that further optimization is required for this platform in the future. Our work shows that silkworm pupae, as an efficient bioreactor, have great potential for application in the production of gene therapy vectors.

scholarly journals Astrocytes Are More Vulnerable than Neurons to Silicon Dioxide Nanoparticle Toxicity in Vitro

Toxics ◽  
2020 ◽  
Vol 8 (3) ◽  
pp. 51
Author(s):  
Jorge Humberto Limón-Pacheco ◽  
Natalie Jiménez-Barrios ◽  
Alejandro Déciga-Alcaraz ◽  
Adriana Martínez-Cuazitl ◽  
Mónica Maribel Mata-Miranda ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Silicon Dioxide ◽  
Culture Media ◽  
Brain Cell ◽  
Attenuated Total Reflection ◽  
Random Coil ◽  
Silicon Dioxide Nanoparticles ◽  
Neurotoxic Effects ◽  
Α Helix

Some studies have shown that silicon dioxide nanoparticles (SiO2-NPs) can reach different regions of the brain and cause toxicity; however, the consequences of SiO2-NPs exposure on the diverse brain cell lineages is limited. We aimed to investigate the neurotoxic effects of SiO2-NP (0–100 µg/mL) on rat astrocyte-rich cultures or neuron-rich cultures using scanning electron microscopy, Attenuated Total Reflection-Fourier Transform Infrared spectroscopy (ATR-FTIR), FTIR microspectroscopy mapping (IQ mapping), and cell viability tests. SiO2-NPs were amorphous particles and aggregated in saline and culture media. Both astrocytes and neurons treated with SiO2-NPs showed alterations in cell morphology and changes in the IR spectral regions corresponding to nucleic acids, proteins, and lipids. The analysis by the second derivative revealed a significant decrease in the signal of the amide I (α-helix, parallel β-strand, and random coil) at the concentration of 10 µg/mL in astrocytes but not in neurons. IQ mapping confirmed changes in nucleic acids, proteins, and lipids in astrocytes; cell death was higher in astrocytes than in neurons (10–100 µg/mL). We conclude that astrocytes were more vulnerable than neurons to SiO2-NPs toxicity. Therefore, the evaluation of human exposure to SiO2-NPs and possible neurotoxic effects must be followed up.

Human insulin gene expressing with Bombyx mori multiple nucleopolyhedrovirus (BmMNPV) expression system

2010 ◽  
Vol 27 (2) ◽  
pp. 393-399
Author(s):  
Wan-fu Yue ◽  
Fang Zhou ◽  
Jia-biao Hu ◽  
Enoch Y. Park ◽  
Joe Hull ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Human Insulin ◽  
Expression System ◽  
Insulin Gene ◽  
Multiple Nucleopolyhedrovirus ◽  
Human Insulin Gene ◽  
Gene Expressing

Construction of a binary transgenic gene expression system for recombinant protein production in the middle silk gland of the silkworm Bombyx mori

2009 ◽  
Vol 19 (3) ◽  
pp. 473-487 ◽  
Author(s):  
Ken-ichiro Tatematsu ◽  
Isao Kobayashi ◽  
Keiro Uchino ◽  
Hideki Sezutsu ◽  
Tetsuya Iizuka ◽  
...  
Keyword(s):  
Gene Expression ◽  
Recombinant Protein ◽  
Bombyx Mori ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Expression System ◽  
Silk Gland ◽  
Gene Expression System ◽  
Silkworm Bombyx Mori

Influence of Post-Treatment with Methanol Vapor on the Properties of SF/P(LLA-CL) Nanofibrous Scaffolds

2011 ◽  
Vol 236-238 ◽  
pp. 2221-2224
Author(s):  
Kui Hua Zhang ◽  
Xiu Mei Mo
Keyword(s):  
Electrospun Nanofibers ◽  
Random Coil ◽  
Methanol Vapor ◽  
Nanofibrous Scaffolds ◽  
Nanofibrous Structure ◽  
Α Helix ◽  
Β Sheet ◽  
Ideal Method ◽  
C Nmr

In order to improve water-resistant ability silk fibroin (SF) and SF/P(LLA-CL) blended nanofibrous scaffolds for tissue engineering applications, methanol vapor were used to treat electrospun nanofibers. SEM indicated SF and SF/ P(LLA-CL) scaffolds maintained nanofibrous structure after treated with methanol vapor and possessed good water-resistant ability. Characterization of 13C NMR clarified methanol vapor induced SF conformation from random coil or α- helix to β-sheet. Moreover, treated SF/ P (LLA-CL) nanofibrous scaffolds still kept good mechanical properties. Methanol vapor could be ideal method to treat SF and SF/ P(LLA-CL) nanofibrous scaffolds for biomedical applications.

Inside Cover: Implantation of Post-translational Tyrosylprotein Sulfation into a Prokaryotic Expression System (ChemBioChem 3/2011)

ChemBioChem ◽  
2011 ◽  
Vol 12 (3) ◽  
pp. 338-338
Author(s):  
Lu-Yi Lu ◽  
Bo-Han Chen ◽  
Jennifer Yun-Shin Wu ◽  
Chen-Chu Wang ◽  
Da-Huang Chen ◽  
...  
Keyword(s):  
Prokaryotic Expression ◽  
Expression System ◽  
Prokaryotic Expression System

α-Helix-to-random-coil transition of two-chain, coiled coils. Theory and experiments for thermal denaturation of α-tropomyosin at acidic pH

1983 ◽  
Vol 16 (3) ◽  
pp. 462-465 ◽  
Author(s):  
Marilyn Emerson Holtzer ◽  
Alfred Holtzer ◽  
Jeffrey Skolnick
Keyword(s):  
Thermal Denaturation ◽  
Coiled Coils ◽  
Random Coil ◽  
Acidic Ph ◽  
Coil Transition ◽  
Α Helix ◽  
Theory And Experiments

scholarly journals Recombinant Dense Granular Protein (GRA5) for Detection of Human Toxoplasmosis by Western Blot

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Xiao Teng Ching ◽  
Yee Ling Lau ◽  
Mun Yik Fong ◽  
Veeranoot Nissapatorn ◽  
Hemah Andiappan
Keyword(s):  
Western Blot ◽  
Affinity Purification ◽  
Expression System ◽  
Economic Loss ◽  
Serum Samples ◽  
Serological Tests ◽  
Human Patient ◽  
Prokaryotic Expression System ◽  
Human Sera ◽  
Lysate Antigens

Toxoplasma gondiiinfects all warm-blooded animals, including humans, causing serious public health problems and great economic loss for the food industry. Commonly used serological tests require costly and hazardous preparation of wholeToxoplasmalysate antigens from tachyzoites. Here, we have evaluated an alternative method for antigen production, which involved a prokaryotic expression system. Specifically, we expressedT. gondiidense granular protein-5 (GRA5) inEscherichia coliand isolated it by affinity purification. The serodiagnostic potential of the purified recombinant GRA5 (rGRA5) was tested through Western blot analysis against 212 human patient serum samples. We found that rGRA5 protein was 100% specific for analysis of toxoplasmosis-negative human sera. Also, rGRA5 was able to detect acute and chronicT. gondiiinfections (sensitivities of 46.8% and 61.2%, resp.).

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