title: [Expression of mouse lipopolysaccharide response protein LRP in E. coli and preparation of rabbit anti-mLRP serum].
abstract: OBJECTIVE
  To express mouse lipopolysaccharide response protein (mLRP) and prepare rabbit anti-mLRP serum.
  
  
  METHODS
  The predicted mouse lrp cDNA sequence was obtained by splicing homologous ESTs by comparing human lrp cDNA with mouse ESTs. Then the primers were designed. mlrp cDNA from NIH3T3 cells stimulated with lipopolysaccharide (LPS) was amplified by RT-PCR and was cloned into prokaryotic expression vector pTAT to construct recombinant expression vector pTAT-mlrp. The His-TAT-mLRP fusion protein was expressed in E. coli BL21(DE3) and was used to immunize the rabbits to get rabbit anti-mLRP serum. The anti-serum was purified by the acetone precipitation method. The specificity of the rabbit anti-mLRP serum was determined by Western blot.
  
  
  RESULTS
  The predicted length of mlrp cDNA was 1905 bp. The encoding region of the cloned mlrp cDNA, 1554 bp, was inserted into pTAT. The His-TAT-mLRP fusion protein was expressed successfully in E. coli. The rabbit anti-mLRP serum was prepared by immunizing the rabbit with mLRP protein.
  
  
  CONCLUSIONS
  The successful expression of mLRP and the preparation of rabbit anti-mLRP serum lays the foundation for further study of the function of mLRP.
authors.affiliation: Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China. daizmly@yahoo.com
authors.forename: Zhong-ming
authors.initials: ZM
authors.lastname: Dai
authors.forename: Su-min
authors.initials: SM
authors.lastname: Chen
authors.forename: Ke-jun
authors.initials: KJ
authors.lastname: Du
authors.forename: Nan-chun
authors.initials: NC
authors.lastname: Chen
authors.forename: Qing-he
authors.initials: QH
authors.lastname: Song
authors.forename: Wen-jing
authors.initials: WJ
authors.lastname: Luo
authors.forename: Jing-yuan
authors.initials: JY
authors.lastname: Chen
mesh_terms: D000209:Acute-Phase Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D000818:Animals; D000917:Antibody Formation / Q000276:immunology*; D002352:Carrier Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D005260:Female; D015870:Gene Expression; D005822:Genetic Vectors; D008070:Lipopolysaccharides / Q000737:chemistry; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D041681:NIH 3T3 Cells; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000099231:Lipopolysaccharide-Binding Protein
chemical_list: D000209:Acute-Phase Proteins; D002352:Carrier Proteins; D008070:Lipopolysaccharides; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; D000099231:Lipopolysaccharide-Binding Protein