title: [Cloning and prokaryotic expression of human secreted IL-16 gene].
abstract: OBJECTIVE
  To clone human secreted IL-16 cDNA, construct its prokaryotic expression vector and express it in E.coli DH5α.
  
  
  METHODS
  The secreted IL-16 gene fragment from the cDNA library of human peripheral blood mononuclear cells (PBMC) was amplified by PCR. After purified, the product was cloned into pUC18 T-vector. The recombinant plasmid was confirmed by PCR, endonuclease digestion and sequencing analysis and then subcloned into prokaryotic expression vector pMAL-C2. Then the recombinant expression plasmid pMAL-IL-16 was transformed into E.coli DH5α. The expression of secreted hIL-16 was induced with IPTG and identified by SDS-PAGE and Western blotting.
  
  
  RESULTS
  Using PCR, we obtained the human secreted IL-16 cDNA fragment which was 393 bp and encoded 130 amino acids. The prokaryotic expression vector pMAL-IL-16 we constructed was successfully transformed into E.coli DH5α, and under the induction of IPTG, we found the expression of the recombinant fusion protein with relative molecular weight (M(r);) being 56 000 as expected and confirmed by SDS-PAGE and Western blotting.
  
  
  CONCLUSIONS
  We successfully cloned the human secreted IL-16 cDNA and constructed its prokaryotic expression vector and expressed it in E.coli, which is helpful for further purification of human secreted IL-16 protein.
authors.affiliation: Department of Immunology, Jining Medical College, Jining 272067, China. chipsi@163.com
authors.forename: Chuan-ping
authors.initials: CP
authors.lastname: Si
authors.forename: Shu-ling
authors.initials: SL
authors.lastname: Li
authors.forename: Jia
authors.initials: J
authors.lastname: Fu
authors.forename: Chun-xia
authors.initials: CX
authors.lastname: Li
authors.forename: Zhi-hua
authors.initials: ZH
authors.lastname: Li
authors.forename: Zhi-gang
authors.initials: ZG
authors.lastname: Song
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D019410:Interleukin-16 / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D019410:Interleukin-16