title: Cloning and expression of special F protein from human liver.
abstract: OBJECTIVE
  To clone human liver special F protein and to express it in a prokaryotic system.
  
  
  METHODS
  Total RNA was isolated from human liver tissue and first-strand cDNA was reverse transcribed using the PCR reverse primer. Following this, cDNA of the F protein was ligated into the clone vector pUCm-T. The segment of F protein's cDNA was subcloned into the expression vector pET-15b and transformed into E. coli BL21 (DE3) pLyss. Isopropy-beta-D-thiogalactoside (IPTG) was then used to induce expression of the target protein.
  
  
  RESULTS
  The cDNA clone of human liver special F protein (1134bp) was successfully produced, with the cDNA sequence being published in Gene-bank: DQ188836. We confirmed the expression of F protein by Western blot with a molecular weight of 43 kDa. The expressed protein accounted for 40% of the total protein extracted.
  
  
  CONCLUSIONS
  F protein expresses cDNA clone in a prokaryotic system, which offers a relatively simple way of producing sufficient quantities of F protein and contributes to understanding the principal biological functions of this protein.
authors.affiliation: Tianjin Third Central Hospital, Tianjin Artificial Cell Laboratory, Tianjin 300170, China. lshye@163.com
authors.forename: Shu-Ye
authors.initials: SY
authors.lastname: Liu
authors.forename: Xin-Da
authors.initials: XD
authors.lastname: Yu
authors.forename: Chun-Juan
authors.initials: CJ
authors.lastname: Song
authors.forename: Wei
authors.initials: W
authors.lastname: Lu
authors.forename: Jian-Dong
authors.initials: JD
authors.lastname: Zhang
authors.forename: Xin-Rong
authors.initials: XR
authors.lastname: Shi
authors.forename: Ying
authors.initials: Y
authors.lastname: Duan
authors.forename: Ju
authors.initials: J
authors.lastname: Zhang
mesh_terms: D003001:Cloning, Molecular / Q000379:methods*; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006801:Humans; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics
chemical_list: D018076:DNA, Complementary; C521996:F protein, human; D011506:Proteins; D012333:RNA, Messenger