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Updated: Aug 11, 2026
A Seamless Cloning Approach for Porcine Reproductive and Respiratory Syndrome Virus Expression Vector Construction
Published on: May 17, 2024
[Cloning and prokaryotic expression of truncated muramidase-released protein gene of human Streptococcus suis type 2]
Hai-li Wang1, Chang-jun Wang, Cheng-ping Lu
1Institute of Military Medical Sciences, Nanjing Command, Nanjing 210002, China. w.haili@163.com
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.
