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[Cloning and prokaryotic expression of truncated muramidase-released protein gene of human Streptococcus suis type 2].

AIM: To clone and express the truncated Habb mrp gene of human Streptococcus suis type 2 (S.suis 2) and detect its activity. METHODS: A pair of primers based on S.suis 2 mrp gene were schemed out. The turncated mrp (tmrp)gene of S.suis 2 strain Habb isolated from diseased person in Haian, Jiangsu province was cloned and analyzed. The prokaryotic expression plasmid pGEX4T-2-tmrp was constructed.The expression of recombinant protein with glutathione S-transferase (GST) was induced in E.coli TG1. The fusion protein (tMRP-GST) was purified by affinity chromatography, and the GST was cut from tMRP-GST with thrombin protease to gain the truncated MRP (tMRP) antigen. The activity of recombinant protein was analyzed by Western blot. RESULTS: Sequence analysis showed that the length of the truncated mrp was 957 bp. The prokaryotic expressed production was a fusion protein, whose molecular weight was about 61 kD, and the molecular weight of the purified tMRP protein was about 35 kD. Western blot analysis showed that tMRP-GST and tMRP were detected specifically by MRP antiserum on nitrocellulose membrane. CONCLUSION: The truncated mrp gene of human S.suis 2 strain Habb is successfully cloned, and the high expression of the functional recombinant protein is achieved in the prokaryotic system, which facilitates the further studies on the bio-function and immunology.

Animals↗

Serum and effluent lysozyme (muramidase) activity in CAPD patients.

The elevated serum lysozyme activity in 13 chronic renal failure patients (n = 26) dropped significantly during their first three months of CAPD and subsequently returned to initial levels. When compared with peritoneal mass transfer of lysozyme and serum creatinine levels, a distinct correlation was observed between these and the fluctuations in serum lysozyme activity recorded up to three years of CAPD (r = 0.319, P less than 0.05 and r = 0.425, P less than 0.025, respectively). A notable drop in the mass transfer of this low molecular weight protein took place after the first hour of dialysis. We concluded that long-term CAPD does not affect serum lysozyme activity and that passive loss across the peritoneal membrane could account for the lysozyme found in the effluent fluid.

Adolescent↗

Lack of correlation between serum and salivary concentration levels of immunoglobulin A and lysozyme (muramidase).

The study was conducted in a group of 232 individuals comprising 114 males and 118 females 20 to 67 years old. As revealed by the linear regression analysis method, mean serum concentration levels of immunoglobulin A (IgA) and lysozyme (LYS) were unrelated to values found in the spontaneously secreted saliva specimens. Levels of serum IgA were clearly age-dependent, but no such correlation was recorded in the case of secretory IgA. These findings suggest that tests for salivary IgA can hardly be taken as an equivalent to IgA determinations in the human serum.

Adult↗

Serum lysozyme (muramidase) levels in healthy subjects aged 10-90 years.

Lysozyme (LYS) levels were tested in the serum of 1,399 subjects (670 males, 729 females). A curve was plotted from the mean values. In our series, both sexes attained their highest peak at about 10 years. Between 21 and 30 years the curves of the two sexes began to diverge and in the 31- to 40-year decade the male curve attained a second maximum. This inter-sex difference was highly significant (P less than 0.01). Between 41 and 50 years the LYS values of the two sexes began to come closer again and from the 51- to 60-year decade they ran parallel with each other without deviating. The significance of the two courses is discussed.

Adolescent↗