scholarly journals Burkholderia thailandensis oacAMutants Facilitate the Expression ofBurkholderia mallei-Like O Polysaccharides

2010 ◽  
Vol 79 (2) ◽  
pp. 961-969 ◽  
Author(s):  
Paul J. Brett ◽  
Mary N. Burtnick ◽  
Christian Heiss ◽  
Parastoo Azadi ◽  
David DeShazer ◽  
...  
Keyword(s):  
Methylation Status ◽  
Cost Effective ◽  
Resonance Spectroscopy ◽  
Acetylation Phenotype ◽  
Burkholderia Thailandensis ◽  
Reading Frame ◽  
Vaccine Candidates ◽  
Identification And Characterization ◽  
Effective Source

ABSTRACTPrevious studies have shown that the O polysaccharides (OPS) expressed byBurkholderia malleiare similar to those produced byBurkholderia thailandensisexcept that they lack the 4-O-acetyl modifications on their 6-deoxy-α-l-talopyranosyl residues. In the present study, we describe the identification and characterization of an open reading frame, designatedoacA, expressed byB. thailandensisthat accounts for this phenomenon. Utilizing theB. thailandensisandB. malleilipopolysaccharide (LPS)-specific monoclonal antibodies Pp-PS-W and 3D11, Western immunoblot analyses demonstrated that the LPS antigens expressed by theoacAmutant,B. thailandensisZT0715, were antigenically similar to those produced byB. malleiATCC 23344. In addition, immunoblot analyses demonstrated that whenB. malleiATCC 23344 was complemented intranswithoacA, it synthesizedB. thailandensis-like LPS antigens. To elucidate the structure of the OPS moieties expressed by ZT0715, purified samples were analyzed via nuclear magnetic resonance spectroscopy. As predicted, these studies demonstrated that the loss of OacA activity influenced the O acetylation phenotype of the OPS moieties. Unexpectedly, however, the results indicated that the O methylation status of the OPS antigens was also affected by the loss of OacA activity. Nonetheless, it was revealed that the LPS moieties expressed by theoacAmutant reacted strongly with theB. malleiLPS-specific protective monoclonal antibody 9C1-2. Based on these findings, it appears that OacA is required for the 4-Oacetylation and 2-Omethylation ofB. thailandensisOPS antigens and that ZT0715 may provide a safe and cost-effective source ofB. mallei-like OPS to facilitate the synthesis of glanders subunit vaccine candidates.

scholarly journals Identification and characterization of antigens as vaccine candidates againstKlebsiella pneumoniae

2013 ◽  
Vol 9 (3) ◽  
pp. 497-505 ◽  
Author(s):  
Urban Lundberg ◽  
Beatrice M. Senn ◽  
Wolfgang Schüler ◽  
Andreas Meinke ◽  
Markus Hanner
Keyword(s):  
Vaccine Candidates ◽  
Identification And Characterization

Cloning and characterization of an alternative splicing transcript of the gene coding for human cytidine deaminase

2007 ◽  
Vol 85 (1) ◽  
pp. 96-102 ◽  
Author(s):  
Bianca Cristina Garcia Lisboa ◽  
Tamara da Rocha Machado ◽  
Daniel Carvalho Pimenta ◽  
Sang Won Han
Keyword(s):  
Alternative Splicing ◽  
Dna Sequences ◽  
Genomic Sequence ◽  
Cytidine Deaminase ◽  
Alternative Form ◽  
Nih3t3 Cells ◽  
Reading Frame ◽  
Gene Coding ◽  
Identification And Characterization

Human cytidine deaminase (HCD) catalyzes the deamination of cytidine or deoxycytidine to uridine or deoxyuridine, respectively. The genomic sequence of HCD is formed by 31 kb with 4 exons and several alternative splicing signals, but an alternative form of HCD has yet to be reported. Here we describe the cloning and characterization of a small form of HCD, HSCD, and it is likely to be a product of alternative splicing of HCD. The alignment of DNA sequences shows that the HSCD matches HCD in 2 parts, except for a deletion of 170 bp. Based on the HCD genome organization, exons 1 and 4 should be joined and all sequences of introns and exons 2 and 3 should be deleted by splicing. This alternative splicing shifted the translation of the reading frame from the point of splicing. The estimated molecular mass is 9.8 kDa, and this value was confirmed by Western blot and mass spectroscopy after expressing the gene fused with glutathionine-S-transferase in the pGEX vector. The deletion and shift of the reading frame caused a loss of HCD activity, which was confirmed by enzyme assay and also with NIH3T3 cells modified to express HSCD and challenged against cytosine arabinoside. In this work we describe the identification and characterization of HSCD, which is the product of alternative splicing of the HCD gene.

Identification and characterization of G90, a novel mouse RNA that lacks an extensive open reading frame

Gene ◽  
1999 ◽  
Vol 232 (1) ◽  
pp. 35-42 ◽  
Author(s):  
Ralf Krause ◽  
Myriam Hemberger ◽  
Heinz Himmelbauer ◽  
Vera Kalscheuer ◽  
Reinald H. Fundele
Keyword(s):  
Open Reading Frame ◽  
Reading Frame ◽  
Identification And Characterization

scholarly journals Yos1p Is a Novel Subunit of the Yip1p–Yif1p Complex and Is Required for Transport between the Endoplasmic Reticulum and the Golgi Complex

2005 ◽  
Vol 16 (4) ◽  
pp. 1673-1683 ◽  
Author(s):  
Matthew Heidtman ◽  
Catherine Z. Chen ◽  
Ruth N. Collins ◽  
Charles Barlowe
Keyword(s):  
Endoplasmic Reticulum ◽  
Golgi Complex ◽  
Secretory Pathway ◽  
Integral Membrane Protein ◽  
Rab Gtpases ◽  
Reading Frame ◽  
Multicopy Suppressor ◽  
Transport Vesicles ◽  
Identification And Characterization

Yeast Yip1p is a member of a conserved family of transmembrane proteins that interact with Rab GTPases. Previous studies also have indicated a role for Yip1p in the biogenesis of endoplasmic reticulum (ER)-derived COPII transport vesicles. In this report, we describe the identification and characterization of the uncharacterized open reading frame YER074W-A as a novel multicopy suppressor of the thermosensitive yip1-4 strain. We have termed this gene Yip One Suppressor 1 (YOS1). Yos1p is essential for growth and for function of the secretory pathway; depletion or inactivation of Yos1p blocks transport between the ER and the Golgi complex. YOS1 encodes an integral membrane protein of 87 amino acids that is conserved in eukaryotes. Yos1p localizes to ER and Golgi membranes and is efficiently packaged into ER-derived COPII transport vesicles. Yos1p associates with Yip1p and Yif1p, indicating Yos1p is a novel subunit of the Yip1p–Yif1p complex.

scholarly journals Identification and Characterization of a Novel Nontranslated Sequence Variant of the Human Intestinal Di-/Tripeptide Transporter, hPEPT1

2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
Helle Bach Søndergaard ◽  
Carsten Uhd Nielsen ◽  
Birger Brodin
Keyword(s):  
Sequence Variant ◽  
Regulatory Effect ◽  
Transport Activity ◽  
Reading Frame ◽  
Translation Analysis ◽  
Nucleotide Insertions ◽  
Identification And Characterization

The human H+-coupled di-/tripeptide transporter (hPEPT1) mediates intestinal absorption of dietary di- and tripeptides, as well as several peptidomimetic drug compounds. The aim of the present study was to investigate the possible role of the hPEPT1 variant hPEPT1-RF in hPEPT1 regulation. However, the proposed hPEPT1-RF mRNA sequence could not be detected in Caco-2 cells or in human intestinal samples. Instead, a new sequence variant, hPEPT1-RFI, was found, which is almost identical to the proposed hPEPT1-RF, except for two nucleotide insertions and one deletion that resulted in a changed open reading frame as compared to hPEPT1-RF. In vitro translation analysis showed that hPEPT1-RFI was not translated. In conclusion, the existence of hPEPT1-RF could not be confirmed; furthermore, the identified sequence variant, hPEPT1-RFI, does not appear to be translated and is therefore unlikely to have a regulatory effect on hPEPT1 transport activity.

Identification and characterization of the gene products of open reading frame U86/87 of human herpesvirus 6

2002 ◽  
Vol 89 (1) ◽  
pp. 89-101 ◽  
Author(s):  
Eleni Papanikolaou ◽  
Vlassis Kouvatsis ◽  
Georgios Dimitriadis ◽  
Naoki Inoue ◽  
Minas Arsenakis
Keyword(s):  
Human Herpesvirus ◽  
Open Reading Frame ◽  
Gene Products ◽  
Human Herpesvirus 6 ◽  
Reading Frame ◽  
Herpesvirus 6 ◽  
Identification And Characterization

scholarly journals Identification and characterization of a novel potyvirus infecting Paris yunnanensis

2021 ◽  
Author(s):  
Pingxiu Lan ◽  
Peng He ◽  
Mengji Cao ◽  
Guohua Zhou ◽  
Li Chenrong ◽  
...  
Keyword(s):  
High Throughput Sequencing ◽  
Genomic Sequence ◽  
Pairwise Comparison ◽  
Distinct Species ◽  
Reading Frame ◽  
Single Clade ◽  
Terminal Poly ◽  
Identification And Characterization ◽  
Complete Genomic

Abstract The complete genomic sequence of a novel potyvirus from Paris yunnanensis was determined by high-throughput sequencing then confirmed by Sanger sequencing. Its genomic RNA consists 9600 nucleotides (nt) excluding the 3’-terminal poly (A) tail, containing a typical large open reading frame (ORF) of potyviruses and encoding a putative polyprotein of 3098 amino acids (aa). Pairwise comparison analysis showed the virus shares sequence identity with other members of Potyvirus was 53.0–57.8% at genome sequence level, and 39.3–51.2% at polyprotein sequence level. Phylogenetic analysis indicated that the virus was clustered as a single clade within the genus Potyvirus both using nt and aa level. These results suggest that the virus should be considered as a distinct species within the genus Potyvirus, and it was tentatively named as “Paris mottle virus” (PaMoV).

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