Past Issues in 2014

Volume: 4 Issue: 1

Biochemistry

Cell Surface Protein-protein Binding on COS-7 Cells Authors: Kae-Jiun Chang and Matthew N. Rasband, date: 01/05/2014, view: 6659, Q&A: 0

Examination of interactions between a transmembrane protein and a soluble protein by pull-down or immunoprecipitation assays can be tricky and complicated due to the detergent extraction of membrane proteins during the lysate preparation step. The choice and concentration of detergents must be determined empirically and the procedure can be ...

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Immunology

T Follicular Helper Cell Coculture Assay Authors: Rafael A. Cubas and Elias K. Haddad, date: 01/05/2014, view: 10480, Q&A: 0

T follicular helper (Tfh) cells constitute a distinct subset of CD4⁺ T cells specialized in providing help to B cells in germinal centers. Phenotypically, Tfh cells are characterized by their high expression of the chemokine receptor CXCR5 that allows their migration into B cell follicles as well as high expression of PD-1, BTLA, the ...

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Microbiology

Detection of piggyBac-mediated Transposition by Splinkerette PCR in Transgenic Lines of Strongyloides ratti Authors: Hongguang Shao and James B. Lok, date: 01/05/2014, view: 9320, Q&A: 0

Splinkerette PCR (spPCR) is a newly developed and efficient method to ascertain and characterize genomic insertion sites of transgenes. The method described in this protocol was successfully applied to confirm piggyBac transposon-mediated integration of transgenes into chromosomes of the parasitic nematode Strongyloides ratti. ...

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Plant Science

In vitro Nematode Infection on Potato Plant Authors: Demosthenis Chronis, Shiyan Chen, Ping Lang, Tien Tran, David Thurston and Xiaohong Wang, date: 01/05/2014, view: 9455, Q&A: 0

Potato cyst nematodes (PCNs; Globodera rostochiensis and G. pallida) are devastating pests that infect potato root. We describe an in vitro assay for PCN infection on potato plantlet in tissue culture. This method is useful for studying nematode parasitism on potato and for investigating responses of potato clones/lines ...

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Potato Transformation Authors: Demosthenis Chronis, Shiyan Chen, Ping Lang, Tien Tran, David Thurston and Xiaohong Wang, date: 01/05/2014, view: 11320, Q&A: 0

This is a protocol to produce stable transgenic potato plants (Solanum tuberosum cv. Désirée) by Agrobacterium-mediated genetic transformation, which is established based on a method described by (Jung et al., 2005) with some modifications. Agrobacterium tumefaciens strain LBA4404 carrying the desired construct ...

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Subcellular Localization Experiments and FRET-FLIM Measurements in Plants Authors: Malgorzata Lichocka and Elmon Schmelzer, date: 01/05/2014, view: 10282, Q&A: 0

Determining the localization of proteins within living cells may be very essential for understanding their biological function. Usually for analysis of subcellular localization, a construct encoding the translational fusion of a cDNA of interest with a fluorescent protein (FP) is engineered, transiently expressed in plant cells and examined with ...

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Biolistic Bombardment for Co-expression of Proteins Fused to YFP and mRFP in Leaf Epidermal Cells of Phaseolus vulgaris ‘Red Mexican’ Author: Malgorzata Lichocka, date: 01/05/2014, view: 8300, Q&A: 0

Biolistic bombardment is based on coating of tungsten or gold particles with DNA and delivery of these “biobullets” into living plant cells under high pressure (Sudowe and Reske-Kunz, 2013). This method enables transient expression of a DNA construct encoding fusion of the protein of interest to a fluorescence protein e.g. GFP for ...

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Agrobacterium-mediated Transformation of Strawberry Author: Jan G. Schaart, date: 01/05/2014, view: 8343, Q&A: 0

Traditional breeding for improvement of strawberry (Fragaria x ananassa) is difficult because strawberry is an octoploid, hybrid species. Genetic modification of strawberry would though be a promising alternative for obtaining the desired improvements in existing elite strawberry cultivars (Schaart et al., 2011). The availability ...

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