title: [Expression of glutathione-S-transferase fusion protein and human CCL3L1 protein].
abstract: OBJECTIVE
  To clone human CCL3L1 cDNA and to express and purify the glutathione-S-transferase (GST) fusion protein and human CCL3L1 protein.
  
  
  METHODS
  Total RNA was isolated from breast cancer cell line MCF7. CCL3L1 cDNA including open reading frame was obtained by RT-PCR. PCR product was digested with EcoR I and cloned into the pGEX-4T-1 vector. The plasmids from positive clone was prepared and sequenced to confirm the CCL3L1 in correct fusion form. pGEX-4T-CCL3L1 was transfected to BL21 E. coli via isopropyl-beta-D-thiogalactoside (IPTG) induction to produce GST-CCL3L1 fusion protein, which was further detected by SDS-PAGE and Western blotting.
  
  
  RESULTS
  As shown and confirmed by restriction endonuclease digestion analysis, CCL3L1 was correctly inserted into pGEX-4T-1 vector. The expressed fusion protein had a relative molecular weight of approximately 34 kD.
  
  
  CONCLUSIONS
  GST-CCL3L1 fusion protein can be successfully expressed using appropriate vector.
authors.affiliation: Department of Infectious Diseases, Beijing Youan Hospital, Capital University of Medical Sciences, Beijing 100069, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Xu
authors.forename: Ying
authors.initials: Y
authors.lastname: Shi
authors.forename: Jun-Hong
authors.initials: JH
authors.lastname: Li
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Hao
authors.initials: H
authors.lastname: Wu
authors.forename: De-xi
authors.initials: DX
authors.lastname: Chen
mesh_terms: D045744:Cell Line, Tumor; D019742:Chemokines, CC / Q000096:biosynthesis* / Q000235:genetics; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D005260:Female; D005822:Genetic Vectors; D005982:Glutathione Transferase / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: C492767:CCL3L1 protein, human; D019742:Chemokines, CC; D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D007544:Isopropyl Thiogalactoside; D005982:Glutathione Transferase