title: [Prokaryotic soluble expression of protein D of Haemophilus influenzae type b].
abstract: OBJECTIVE
  To express the recombinant D protein in prokaryotic expression system solubly and make preparation for producing D-carrier conjugate vaccine next step.
  
  
  METHODS
  The 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.
  
  
  RESULTS
  The 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.
  
  
  CONCLUSIONS
  The D protein recombined expression plasmid was constructed successfully and expressed D protein in prokaryotic cells in a solube form.
authors.affiliation: Chinese Academy of Medical Sciences, Peking Union Medical College, Institute of Medical Biology, Kunming 650118, China.
authors.forename: Meng-Meng
authors.initials: MM
authors.lastname: Yin
authors.forename: Qiu-Dong
authors.initials: QD
authors.lastname: Su
authors.forename: Min-Zhuo
authors.initials: MZ
authors.lastname: Gyo
authors.forename: Yi-Na
authors.initials: YN
authors.lastname: Cun
authors.forename: Yuan-Qian
authors.initials: YQ
authors.lastname: Pu
authors.forename: Zhi-Yuan
authors.initials: ZY
authors.lastname: Jia
authors.forename: Jing-Ran
authors.initials: JR
authors.lastname: Yang
authors.forename: Yang
authors.initials: Y
authors.lastname: Tang
authors.forename: Guo-Yang
authors.initials: GY
authors.lastname: Liao
authors.forename: Yao
authors.initials: Y
authors.lastname: Yi
authors.forename: Sheng-Li
authors.initials: SL
authors.lastname: Bi
authors.forename: Wei-Dong
authors.initials: WD
authors.lastname: Li
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D015153:Blotting, Western; D002352:Carrier Proteins / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D019741:Haemophilus influenzae type b / Q000235:genetics*; D007072:Immunoglobulin D / Q000235:genetics*; D008074:Lipoproteins / Q000235:genetics*; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis; D012995:Solubility
chemical_list: D001426:Bacterial Proteins; D002352:Carrier Proteins; D007072:Immunoglobulin D; D008074:Lipoproteins; D011994:Recombinant Proteins; C067989:glpQ protein, Haemophilus influenzae