1.Prokaryotic soluble expression of protein D of Haemophilus influenzae type b.
Meng-Meng YIN ; Qiu-Dong SU ; Min-Zhuo GYO ; Yi-Na CUN ; Yuan-Qian PU ; Zhi-Yuan JIA ; Jing-Ran YANG ; Yang TANG ; Guo-Yang LIAO ; Yao YI ; Sheng-Li BI ; Wei-Dong LI
Chinese Journal of Experimental and Clinical Virology 2013;27(2):89-91
OBJECTIVETo express the recombinant D protein in prokaryotic expression system solubly and make preparation for producing D-carrier conjugate vaccine next step.
METHODSThe hpd gene fragment removed of signal peptide from genomic DNA of Hib CMCC was inserted into pET43. 1a. The recombinant plasmid was transformed to competent E. coli BL21 (DE3) for expression under induction of IPTG. The expressed recombination protein was precipitated with ammonium sulfate, purified by DEAE anion exchange column chromatography and identified for reactogenicity by Western Blot.
RESULTSThe expressed recombination protein, in a soluble form, constained about 50% of total somatic protein and showed specific reaction with the HIB antisera after preliminary purification.
CONCLUSIONThe D protein recombined expression plasmid was constructed successfully and expressed D protein in prokaryotic cells in a solube form.
Bacterial Proteins ; genetics ; Blotting, Western ; Carrier Proteins ; genetics ; Escherichia coli ; genetics ; Haemophilus influenzae type b ; genetics ; Immunoglobulin D ; genetics ; Lipoproteins ; genetics ; Plasmids ; Recombinant Proteins ; biosynthesis ; Solubility
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