scholarly journals Enhanced Stability of DNA Oligonucleotides with Partially Zwitterionic Backbone Structures in Biological Media

Molecules ◽  
2018 ◽  
Vol 23 (11) ◽  
pp. 2941 ◽  
Author(s):  
Melissa Meng ◽  
Boris Schmidtgall ◽  
Christian Ducho
Keyword(s):  
Modified Oligonucleotides ◽  
Biological Media ◽  
Whole Cell Lysate ◽  
Cell Lysate ◽  
Dna Oligonucleotides ◽  
Dna Strands ◽  
Backbone Modification ◽  
Backbone Structure ◽  
Oligonucleotide Analogues ◽  
Enhanced Stability

Deficient stability towards nuclease-mediated degradation is one of the most relevant tasks in the development of oligonucleotide-derived biomedical agents. This hurdle can be overcome through modifications to the native oligonucleotide backbone structure, with the goal of simultaneously retaining the unique hybridization properties of nucleic acids. The nucleosyl amino acid (NAA)-modification is a recently introduced artificial cationic backbone linkage. Partially zwitterionic NAA-modified oligonucleotides had previously shown hybridization with DNA strands with retained base-pairing fidelity. In this study, we report the significantly enhanced stability of NAA-modified oligonucleotides towards 3′- and 5′-exonuclease-mediated degradation as well as in complex biological media such as human plasma and whole cell lysate. This demonstrates the potential versatility of the NAA-motif as a backbone modification for the development of biomedically active oligonucleotide analogues.

scholarly journals Nano-LC FTICR Tandem Mass Spectrometry for Top-Down Proteomics: Routine Baseline Unit Mass Resolution of Whole Cell Lysate Proteins up to 72 kDa

2012 ◽  
Vol 84 (5) ◽  
pp. 2111-2117 ◽  
Author(s):  
Jeremiah D. Tipton ◽  
John C. Tran ◽  
Adam D. Catherman ◽  
Dorothy R. Ahlf ◽  
Kenneth R. Durbin ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Tandem Mass Spectrometry ◽  
Mass Resolution ◽  
Unit Mass ◽  
Tandem Mass ◽  
Top Down ◽  
Whole Cell ◽  
Whole Cell Lysate ◽  
Cell Lysate ◽  
Unit Mass Resolution

scholarly journals Effect of Different Adjuvants on Protection and Side-Effects Induced by Helicobacter suis Whole-Cell Lysate Vaccination

PLoS ONE ◽  
2015 ◽  
Vol 10 (6) ◽  
pp. e0131364 ◽  
Author(s):  
Iris Bosschem ◽  
Jagadeesh Bayry ◽  
Ellen De Bruyne ◽  
Kim Van Deun ◽  
Annemieke Smet ◽  
...  
Keyword(s):  
Side Effects ◽  
Whole Cell ◽  
Whole Cell Lysate ◽  
Cell Lysate

Proteomics workflow for whole cell lysate, endosome, and lysosome fractions v2

2021 ◽  
Author(s):  
Hankum Park ◽  
Frances V Hundley ◽  
J. Wade Harper
Keyword(s):  
Sample Preparation ◽  
Whole Cell ◽  
Cell Lysates ◽  
Whole Cell Lysate ◽  
Cell Lysate ◽  
Ms Analysis

We present a protocol for sample preparation for LC-MS analysis of whole cell lysates and for lysosomal and endosomal fractions purified by Lyso-IP and Endo-IP. Protocols for purification of lysosomes and endosomes is provided in protocol dx.doi.org/10.17504/protocols.io.byi9puh6 using cells that express endogenously tagged TMEM192-HA and stably expressing FLAG-EEA1 as descrbed in dx.doi.org/10.17504/protocols.io.byi7puhn.

Initiation by RNA polymerase II and formation of runoff transcripts containing unblocked and unmethylated 5' termini

1983 ◽  
Vol 3 (12) ◽  
pp. 2172-2179
Author(s):  
H Ernst ◽  
W Filipowicz ◽  
A J Shatkin
Keyword(s):  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Atp Hydrolysis ◽  
Beta Globin ◽  
Whole Cell ◽  
Whole Cell Lysate ◽  
Cell Lysate ◽  
Promoter Dna ◽  
Gamma S ◽  
Made In

Transcription of cloned adenovirus, beta-globin, and retrovirus long terminal repeat DNAs in HeLa whole-cell lysate was inhibited by S-adenosylhomocysteine. However, full-length 1.7-kilobase transcripts made on adenovirus 2 late promoter DNA contained 5'-terminal GpppA, consistent with specific initiation and runoff synthesis in the absence of product methylation. Formation of runoff transcripts including retrovirus RNAs that normally contain 5'-m7GpppGmpC was not decreased by replacing GTP with non-hydrolyzable analogs, and Rous-associated virus-2 runoff products made in the presence of GTP-gamma-S contained 5'-terminal gamma-S-pppGpC. The results indicate that capping and specific transcript synthesis by RNA polymerase II are not obligatorily linked in HeLa whole-cell lysate. Accurate initiation is dependent on ATP hydrolysis, and in contrast to GTP, replacement of ATP by 5'-adenylyl-imidodiphosphate blocked specific initiation of transcripts that start with either GTP (Rous-associated virus-2, Rous-associated virus-0) or ATP (beta-globin, adenovirus).

Production and preliminary evaluation of Trypanosoma evansi HSP70 for antibody detection in Equids

2015 ◽  
Vol 60 (4) ◽  
Author(s):  
Jaideep Kumar ◽  
Ashok Chaudhury ◽  
Bidhan C. Bera ◽  
Ritesh Kumar ◽  
Rajender Kumar ◽  
...  
Keyword(s):  
Ethical Issues ◽  
Terminal Region ◽  
Antibody Detection ◽  
Preliminary Evaluation ◽  
Large Set ◽  
In Vitro Production ◽  
Serum Samples ◽  
Whole Cell ◽  
Whole Cell Lysate ◽  
Cell Lysate

AbstractThe present immuno-diagnostic method using soluble antigens from whole cell lysate antigen for trypanosomosis have certain inherent problems like lack of standardized and reproducible antigens, as well as ethical issues due to in vivo production, that could be alleviated by in vitro production. In the present study we have identified heat shock protein 70 (HSP70) from T. evansi proteome. The nucleotide sequence of T. evansi HSP70 was 2116 bp, which encodes 690 amino acid residues. The phylogenetic analysis of T. evansi HSP70 showed that T. evansi occurred within Trypanosoma clade and is most closely related to T. brucei brucei and T. brucei gambiense, whereas T. congolense HSP70 laid in separate clade. The two partial HSP70 sequences (HSP-1 from N-terminal region and HSP-2 from C-terminal region) were expressed and evaluated as diagnostic antigens using experimentally infected equine serum samples. Both recombinant proteins detected antibody in immunoblot using serum samples from experimental infected donkeys with T. evansi. Recombinant HSP-2 showed comparable antibody response to Whole cell lysate (WCL) antigen in immunoblot and ELISA. The initial results indicated that HSP70 has potential to detect the T. evansi infection and needs further validation on large set of equine serum samples.

Two-dimensional blue native/SDS-PAGE analysis of whole cell lysate protein complexes of rice in response to salt stress

2016 ◽  
Vol 200 ◽  
pp. 90-101 ◽  
Author(s):  
Amenehsadat Hashemi ◽  
Javad Gharechahi ◽  
Ghorbanali Nematzadeh ◽  
Faezeh Shekari ◽  
Seyed Abdollah Hosseini ◽  
...  
Keyword(s):  
Salt Stress ◽  
Protein Complexes ◽  
Two Dimensional ◽  
Whole Cell ◽  
Whole Cell Lysate ◽  
Cell Lysate ◽  
Sds Page ◽  
Blue Native

scholarly journals ESCORTing proteins directly from whole cell-lysate for single-molecule studies

2017 ◽  
Vol 535 ◽  
pp. 35-42 ◽  
Author(s):  
Shwetha Srinivasan ◽  
Jagadish P. Hazra ◽  
Gayathri S. Singaraju ◽  
Debadutta Deb ◽  
Sabyasachi Rakshit
Keyword(s):  
Single Molecule ◽  
Whole Cell ◽  
Whole Cell Lysate ◽  
Cell Lysate ◽  
Single Molecule Studies
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