Heterologous expression and self-assembly of the S-layer protein SbsA of Bacillus stearothermophilus in Escherichia coli.

Abstract:

:The cell surface of Bacillus stearothermophilus PV72 is covered by a regular surface layer (S-layer) composed of single species of protein, SbsA, with a molecular weight of 130,000. Recently, the sequence of the corresponding gene (sbsA) has been determined. The SbsA coding region including the signal sequence was cloned as a polymerase chain reaction (PCR) product into a low-copy-number vector under the transcriptional control of the lambda pL promoter. Expression of sbsA was shown to be thermally inducible from the resulting vector pBK4 in a strain of Escherichia coli expressing the lambda cl857 from the chromosome. As shown by ultrathin sectioning of whole cells and immunogold labelling using SbsA- specific antibodies, expression of sbsA in E. coli led to accumulation of sheet-like self-assembling products of the protein in the cytoplasm. No SbsA protein was detected either in the periplasm or in the supernatant fractions. Long-term expression of sbsA from pBK4, including in the late stationary phase, did not lead to degradation of SbsA.

journal_name

Mol Microbiol

journal_title

Molecular microbiology

authors

Kuen B,Sára M,Lubitz W

doi

10.1046/j.1365-2958.1996.386918.x

subject

Has Abstract

pub_date

1996-02-01 00:00:00

pages

495-503

issue

3

eissn

0950-382X

issn

1365-2958

journal_volume

19

pub_type

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