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131.
B Guss K Leander U Hellman M Uhlén J Sj?quist M Lindberg 《European journal of biochemistry》1985,153(3):579-585
The protein A (spa) genes from Staphylococcus aureus Cowan I and a mutant strain of Cowan I called V-1 earlier suggested to produce a monovalent IgG-binding protein A have been cloned in Escherichia coli. The DNA sequences coding for the IgG-binding part of the spa genes from both strains have been determined and compared with each other and with a partial amino acid sequence of purified protein A from strain V-1. The nucleotide sequence of the spa gene from strain V-1 reveals an NH2-terminally located IgG-binding region homologous to region E first reported for strain 8325-4, region D and the major portion of region A. The amino acid sequence analysis of the purified protein A from this strain also shows the presence of regions E and D but only a minor part of region A. Reversed-phase high-performance liquid chromatography fractionation of purified protein A from strain V-1 revealed that the preparation was heterogeneous, containing mainly two peptides with different abilities to bind IgG molecules. A shuttle vector containing the cloned protein A gene from V-1 was constructed and transformed into different strains of S. aureus and the produced protein A was purified and analysed using sodium dodecyl sulfate/polyacrylamide gel electrophoresis. 相似文献
132.
133.
Biotechnology Letters - Functional characterization of metagenomic DNA often involves expressing heterologous DNA in genetically tractable microorganisms such as Escherichia coli. Functional... 相似文献
134.
Christopher von Beek Ida Waern Jens Eriksson Fabio Rabelo Melo Carl Robinson Andrew S. Waller Mikael E. Sellin Bengt Guss Gunnar Pejler 《Cellular microbiology》2019,21(9)
Mast cells are implicated in the innate proinflammatory immune defence against bacterial insult, but the mechanisms through which mast cells respond to bacterial encounter are poorly defined. Here, we addressed this issue and show that mast cells respond vividly to wild type Streptococcus equi by up‐regulating a panel of proinflammatory genes and by secreting proinflammatory cytokines. However, this response was completely abrogated when the bacteria lacked expression of sagA, whereas the lack of a range of other potential virulence genes (seeH, seeI, seeL, seeM, hasA, seM, aroB, pyrC, and recA) had no effect on the amplitude of the mast cell responses. The sagA gene encodes streptolysin S, a lytic toxin, and we next showed that the wild type strain but not a sagA‐deficient mutant induced lysis of mast cells. To investigate whether host cell membrane perturbation per se could play a role in the activation of the proinflammatory response, we evaluated the effects of detergent‐ and pneumolysin‐dependent lysis on mast cells. Indeed, exposure of mast cells to sublytic concentrations of all these agents resulted in cytokine responses of similar amplitudes as those caused by wild type streptococci. This suggests that sublytic membrane perturbation is sufficient to trigger full‐blown proinflammatory signalling in mast cells. Subsequent analysis showed that the p38 and Erk1/2 signalling pathways had central roles in the proinflammatory response of mast cells challenged by either sagA‐expressing streptococci or detergent. Altogether, these findings suggest that sagA‐dependent mast cell membrane perturbation is a mechanism capable of activating the innate immune response upon bacterial challenge. 相似文献
135.
Janet L. Guss Harry R. Kissileff Michael J. Devlin Ellen Zimmerli B. Timothy Walsh 《Obesity (Silver Spring, Md.)》2002,10(10):1021-1029
Objective: To determine whether meal size is related to body mass index (BMI) in obese subjects with binge-eating disorder (BED). Research Methods and Procedures: Five groups of subjects each consumed two laboratory-test meals on nonconsecutive days. Forty-two women, categorized by BMI and BED diagnosis, were instructed to “binge” during one meal and to eat “normally” during another. Eighteen women had BMI values >38 kg/m2 (more-obese) and 17 had BMI values between 28 to 32 kg/m2 (less-obese). Twelve of the more-obese and nine of the less-obese individuals met Diagnostic and Statistical Manual (DSM)-IV criteria for BED. Seven normal-weight women also participated as controls. Results: Subjects with BED ate significantly more in both meals than subjects without BED. Binge meals were significantly larger than normal meals only among subjects with BED. The more-obese subjects with BED ate significantly more than the less-obese subjects with BED, but only when they were asked to binge. Intake of the binge meal was significantly, positively correlated with BMI among subjects with BED. Subjects with BED reported significantly higher satiety ratings after the binge than after the normal meal, but subjects without BED reported similar ratings after both meals. Regardless of instructions and diagnosis, obese subjects consumed a significantly higher percentage of energy from fat (38.5%) than did normal-weight subjects (30.8%). Discussion: During binge meals, the energy intake of subjects with BED is greater than that of individuals of similar body weight without BED and is positively correlated with BMI. 相似文献