Protein extraction from the earthworm <i>Eisenia fetida</i> for 2‐DE

Wang, Xing; Chang, Li; Wang, Gaochan; Sun, Zhenjun; Ma, Hongxiang; Sun, Qian; Li, Jing

doi:10.1002/pmic.200900488

Cited by 16 publications

(12 citation statements)

References 24 publications

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“…In turfgrass plants, Xu et al (2008) demonstrated that TCA/ acetone method is reliable method for protein extraction for leaf tissues, while for root tissues, both TCA/acetone and phenol methods can be used. Protein extraction with TCA/acetone base is one of the frequent protocols used to extract proteins from various samples, including plants and insects (Cilia et al 2009;Wang et al 2010;Xie et al 2011;Xu et al 2008;Yin et al 2017;Zhang et al 2015Zhang et al , 2013. TCA/acetone method is an effective method to minimize degradation of protein and to eliminate interfering substances, such as salt and polyphenols (Carpentier et al 2005;Gόmez-Vidal et al 2008;Görg et al 2004).…”

Section: Discussionmentioning

confidence: 99%

“…Numerous protein extraction methods for 1-DE and 2-DE have been developed and applied for various plant samples (Alam et al 2012;Chatterjee et al 2012;Gόmez-Vidal et al 2008;Isaacson et al 2006;Rasool et al 2014;Vincent et al 2006;Xu et al 2008;Zhang et al 2015;Zheng et al 2007). Compared to the optimal methods for the extraction of plant tissues, only a few studies in the literature that compare different extraction methods for insect tissues (Cilia et al 2009;Collet & Baguñà 1993;Qin et al 2013;Wang et al 2010;Yiou et al 2013), particularly for larva and adult tissues of P. xylostella. The major drawback in proteomics research of P. xylostella is the lack of reliable protein extraction methods (Dong et al 2013;Li et al 2014a;Xie et al 2011;Yin et al 2017;Zhang et al 2013).…”

Section: Introductionmentioning

confidence: 99%

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Optimization Method for Proteomic Analysis of the Larva and Adult Tissues of Plutella xylostella (L.) (Lepidoptera: Plutellidae)

Hassan

Yusoff

Aizat

et al. 2018

JSM

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Plutella xylostella (L.) (Lepidoptera: Plutellidae), the major insect pest of cruciferous crops worldwide shows significant resistance to almost all classes of insecticides. In order to effectively prevent and manage the insecticidal resistance, it is crucial to understand the physiological adaptation of insects against insecticides. Identification of insect protein that interacting with insecticides and characterization of their modification in resistant strains can be done by using differential proteomics approach. This study focuses on optimizing a sensitive and rapid method for the extraction of high quality protein of both larva and adult tissues of P. xylostella to be used in two-dimensional gel electrophoresis. Five extraction methods were evaluated for protein concentration, yields and resolving patterns of one-dimensional and two-dimensional electrophoresis. The results showed that trichloroacetic acid/acetone extraction methods with two different concentrations of 2-mercaptoethanol produced the highest protein concentration and yield for both adult and larva tissues, respectively. Meanwhile, trichloroacetic acid/acetone with dithiothreitol extraction method gave better separation of spots and intensity for both larva and adult tissues compared to other methods tested. As such, we concluded that trichloroacetic acid/acetone with dithiothreitol successfully yielded high total protein concentration and good separation of two-dimensional electrophoresis gel spots in both adult and larva P. xylostella.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Optimization Method for Proteomic Analysis of the Larva and Adult Tissues of Plutella xylostella (L.) (Lepidoptera: Plutellidae)

Hassan

Yusoff

Aizat

et al. 2018

JSM

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“…Proteins of a large number of nematodes were extracted according to the trichloroacetic acid (TCA)-acetone procedure described previously [54]. Briefly, nematodes were ground in a mortar with liquid nitrogen until a fine white powder was produced, and then suspended with 3 mL/g fresh weight ice-cold acetone containing 10% TCA and 0.07% dithiothreitol (DTT).…”

Section: Methodsmentioning

confidence: 99%

Screening and Functional Analysis of the Peroxiredoxin Specifically Expressed in Bursaphelenchus xylophilus—The Causative Agent of Pine Wilt Disease

Ren

Huang

et al. 2014

IJMS

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The pine wood nematode, Bursaphelenchus xylophilus, is the causal agent of pine wilt disease. Accurately differentiating B. xylophilus from other nematodes species, especially its related species B. mucronatus, is important for pine wood nematode detection. Thus, we attempted to identify a specific protein in the pine wood nematode using proteomics technology. Here, we compared the proteomes of B. xylophilus and B. mucronatus using Two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization-time-of-flight/time-of-flight (MALDI-TOF/TOF-MS) technologies. In total, 15 highly expressed proteins were identified in B. xylophilus compared with B. mucronatus. Subsequently, the specificity of the proteins identified was confirmed by PCR using the genomic DNA of other nematode species. Finally, a gene encoding a specific protein (Bx-Prx) was obtained. This gene was cloned and expressed in E. coli. The in situ hybridisation pattern of Bx-Prx showed that it was expressed strongly in the tail of B. xylophilus. RNAi was used to assess the function of Bx-Prx, the results indicated that the gene was associated with the reproduction and pathogenicity of B. xylophilus. This discovery provides fundamental information for identifying B. xylophilus via a molecular approach. Moreover, the purified recombinant protein has potential as a candidate diagnostic antigen of pine wilt disease, which may lead to a new immunological detection method for the pine wood nematode.

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“…Proteins were extracted according to the trichloroacetic acid‐A (TCA‐A) procedure described previously 19. Briefly, fresh whole E. fetida samples (2 g) collected from three replications were pulverized to a fine powder with liquid nitrogen and a mortar and pestle.…”

Section: Methodsmentioning

confidence: 99%

Analysis of earthworm Eisenia fetida proteomes during cadmium exposure: An ecotoxicoproteomics approach

et al. 2010

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Identification of differential proteomic responses to cadmium (Cd) would provide a means for better understanding the survival mechanisms of the earthworm, Eisenia fetida, living in a Cd-polluted environment, and the stress responses can be assessed by ecotoxiptoromics approaches. Extracts of whole earthworm collected at days 7, 14, 21, and 28 after Cd exposure were analyzed by 2-DE and quantitative image analysis. In total, 143 proteins demonstrated significant regulation in at least at one time point, and 56 proteins were identified by MALDI-TOF/TOF-MS and database searching. Compared with control samples, 28 protein spots were up-regulated and 28 were down-regulated during at least one time point. The identified proteins, including chaperonine protein HSP60, caspase-8, calcium ion-binding protein, zinc ion-binding protein, actin-binding protein, proteolysis, fibrinolytic protease, glutamate dehydrogenase, gelsolin-like protein, lombricine kinase, coelomic cytolytic factor 1, manganous superoxide dismutase, extracellular globin-4, lysenin, intermediate filament protein, and tubulin, are involved in several processes, including transcription, translation, the tricarboxylic acid cycle, the cellular amino acid metabolic process, protein amino acid phosphorylation, glycolysis, and the glucose metabolic process. Thus, our study provides a functional profile of the Cd-responsive proteins in earthworms.

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Protein extraction from the earthworm Eisenia fetida for 2‐DE

Cited by 16 publications

References 24 publications

Optimization Method for Proteomic Analysis of the Larva and Adult Tissues of Plutella xylostella (L.) (Lepidoptera: Plutellidae)

Optimization Method for Proteomic Analysis of the Larva and Adult Tissues of Plutella xylostella (L.) (Lepidoptera: Plutellidae)

Screening and Functional Analysis of the Peroxiredoxin Specifically Expressed in Bursaphelenchus xylophilus—The Causative Agent of Pine Wilt Disease

Analysis of earthworm Eisenia fetida proteomes during cadmium exposure: An ecotoxicoproteomics approach

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