Expression of the functional recombinant interleukin-16 in E. coli and mammalian cell lines

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Title
Expression of the functional recombinant interleukin-16 in E. coli and mammalian cell lines
Author(s)
Seon-Young Kim; Chang Hun Lee; Kyung Joo Kim; Yeon Soo Kim
Bibliographic Citation
Journal of Microbiology and Biotechnology, vol. 11, no. 2, pp. 234-241
Publication Year
2001
Abstract
The C-terminal 393 bp region of the human interleukin-16 (IL-16) gene was cloned and expressed in E. coli along with mammalian cell lines. Recombinant IL-16 expressed from E. coli was 22 kDa on SDS-PAGE and showed 260% of chemoattractant activity at a concentration of 0.1 μg/ml. HeLa, COS, and Neuro-2a cells were transduced by recombinant retrovirus vector pLNC/IL-16/IRES/TK and the intracellular and secreted amounts of IL-16 produced by HeLa/IL-16/TK, COS/IL-16/TK, and Neuro-2a/IL-16/TK cells were determined by enzyme-linked immunosorbent assay (ELISA). HeLa/IL-16/TK (1 × 105) and COS/IL-16/TK (1 × 105) cells secreted 36.1 and 13.3 ng of IL-16 for 48 h, respectively. Forty-nine ng and 86.4 ng of IL-16 remained in the cell lysates of HeLa/IL-16/TK and COS/IL-16/TK. Intracellular and secreted amounts of IL-16 from Neuro-2a/IL-16/TK (5 × 105) cells during 24 h cultivation were 50 ng and 3.3 ng, respectively. Also, HeLa and COS cells were stably transfected with mammalian expression vector pCRIII/IL-16. Both culture media and cell lysates prepared from HeLa/IL-16 cells and COS/IL-16 cells showed chemoattractant activity ranging from 190% to 460% as compared to the control experiment. Expression of the herpes simplex virus thymidine kinase (HSV-tk) gene in pLNC/IL-16/IRES/TK bicistronic retroviral expression vector was verified by performing a ganciclovir (GCV) sensitivity assay. Finally, IL-16 repressed Tat-transactivated human immunodeficiency virus type 1 long terminal repeat (HIV-1 LTR) promoter activity.
Keyword
E. colimammalian cellsretrovirus producer cellslymphocyte migration activityinterleukin 16HIV-1 LTR suppression
ISSN
1017-7825
Publisher
Korea Soc-Assoc-Inst
Type
Article
Appears in Collections:
1. Journal Articles > Journal Articles
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