A plasmid system for high-level expression and in vitro processing of recombinant proteins

Pohlner, Johannes; Krämer, Joachim; Meyer, Thomas F.

doi:10.1016/0378-1119(93)90354-6

Cited by 36 publications

(26 citation statements)

References 24 publications

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“…When the hinge sequence of IgA1 was substituted for the linker connecting the two domains of endoglucanase A, the fusion protein was cleaved very slowly. Additionally, oligopeptides similar to the IgA1 hinge are cleaved much less efficiently than intact IgA1 (14,19,26,38). Here, using IgA1/IgG2-domain-exchanged proteins, we found that the H. influenzae and N. gonorrhoeae proteases cleave antibodies containing both C ␣ 2 and C ␣ 3, but not antibodies containing C ␣ 1, and lacking either C ␣ 2 or C ␣ 3.…”

Section: Discussionmentioning

confidence: 99%

Cleavage of the Human Immunoglobulin A1 (IgA1) Hinge Region by IgA1 Proteases Requires Structures in the Fc region of IgA

et al. 2003

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Section: Discussionmentioning

confidence: 99%

Cleavage of the Human Immunoglobulin A1 (IgA1) Hinge Region by IgA1 Proteases Requires Structures in the Fc region of IgA

et al. 2003

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“…H. pylori cells were grown for 1 to 3 days (depending on the growth characteristics of the respective strain) in a microaerobic atmosphere (85% N 2 , 10% CO 2 , and 5% O 2 ) at 37°C on serum plates as previously described (31). Escherichia coli strains DH5␣ (BRL) and 2136 (37) were grown on Luria-Bertani (LB) agar plates or in LB liquid medium supplemented with ampicillin (100 mg/liter), if appropriate.…”

Section: Methodsmentioning

confidence: 99%

“…For the generation of an HP0227-specific antiserum, a fusion protein with an N-terminal MS2 polymerase and His tag was produced using the E. coli expression vector pEV40 (37). A fragment of the hp0227 gene (bp 195 to 1107) was amplified by PCR with the primers SO100 (5Ј-GAGAATTCTTCCTTAGTCA ATTTAGCCA-3Ј) and SO101 (5Ј-GACACTCGAGTCAAATGCTGTGGAAA TTGTTC-3Ј), with H. pylori 26695 chromosomal DNA as the template.…”

Section: Methodsmentioning

confidence: 99%

Outer Membrane Protein Expression Profile in Helicobacter pylori Clinical Isolates

et al. 2009

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The gram-negative gastric pathogen Helicobacter pylori is equipped with an extraordinarily large set of outer membrane proteins (OMPs), whose role in the infection process is not well understood. The Hop (Helicobacter outer membrane porins) and Hor (Hop-related proteins) groups constitute a large paralogous family consisting of 33 members. The OMPs AlpA, AlpB, BabA, SabA, and HopZ have been identified as adhesins or adherenceassociated proteins. To better understand the relevance of these and other OMPs during infection, we analyzed the expression of eight different omp genes (alpA, alpB, babA, babB, babC, sabA, hopM, and oipA) in a set of 200 patient isolates, mostly from symptomatic children or young adults. Virtually all clinical isolates produced the AlpA and AlpB proteins, supporting their essential function. All other OMPs were produced at extremely variable rates, ranging from 35% to 73%, indicating a function in close adaptation to the individual host or gastric niche. In 11% of the isolates, BabA was produced, and SabA was produced in 5% of the isolates, but the strains failed to bind their cognate substrates. Interleukin-8 (IL-8) expression in gastric cells was strictly dependent on the presence of the cag pathogenicity island, whereas the presence of OipA clearly enhanced IL-8 production. The presence of the translocated effector protein CagA correlated well with BabA and OipA production. In conclusion, we found unexpectedly diverse omp expression profiles in individual H. pylori strains and hypothesize that this reflects the selective pressure for adhesion, which may differ across different hosts as well as within an individual over time.

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“…Recombinant OipA antigen was produced as a fusion with an N-terminal MS2-polymerase and a His tag by using the Escherichia coli expression vector pEV40 (10). A selected portion of the oipA gene (45 to 882 bp) was PCR amplified with the primers SO102 (5Ј-GAGAATTCCACGCTGAAAGGAA TGGAT-3Ј) and SO103 (5Ј-GATCCTCGAGTCAATAAAC GCTCACCACTCTTT-3Ј) and H. pylori 26695 chromosomal DNA as a template.…”

mentioning

confidence: 99%

Correlation betweenHelicobacter pyloriOipA Protein Expression andoipAGene Switch Status

Kudo

Nurgalieva²,

Conner

et al. 2004

J Clin Microbiol

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A plasmid system for high-level expression and in vitro processing of recombinant proteins

Cited by 36 publications

References 24 publications

Cleavage of the Human Immunoglobulin A1 (IgA1) Hinge Region by IgA1 Proteases Requires Structures in the Fc region of IgA

Cleavage of the Human Immunoglobulin A1 (IgA1) Hinge Region by IgA1 Proteases Requires Structures in the Fc region of IgA

Outer Membrane Protein Expression Profile in Helicobacter pylori Clinical Isolates

Correlation betweenHelicobacter pyloriOipA Protein Expression andoipAGene Switch Status

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