Abstract
Human β-defensin (HBD)-2 is a small cationic peptide with a broad range of antimicrobial activity. In this study, multiple copies of the hBD2 gene were linked in tandem, and a number of different Escherichia coli expression vectors were evaluated, including pQE-30, pBV220, pET-28a(+), and pGEX-4T-2. No expression of multiple joined genes was detectable in the pQE-30 expression system, whereas in pBV220 with one or two joined hBD2 genes and in pET-28a(+) with one, two, or four copies, target proteins were expressed at a low level. Only when pGEX-4T-2 was applied as expression plasmid with one or two joined hBD2 genes were target proteins expressed in high level, and the expressed fusion proteins account for 26 and 16% of the total insoluble proteins, respectively. In the pGEX-4T-2 and pET-28a(+) expression systems, the effects of multiple joined genes on the growth of host strains and plasmid stability were examined. Host cells containing plasmid carrying fewer copies of hBD2 genes were faster in cell growth. Plasmid stability decreased with an increase in multiple joined genes, which was especially noticeable in the pET-28a(+) system. Furthermore, the presence of glucose in culture medium brought about a positive effect on plasmid stability when using pET28-nhBD2 as expression vectors.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Piers, K. L., Brown, M. H., and Hancock, R. E. W. (1993), Gene 134, 7–13.
Lennick, M., Haynes, J. R., and Shen, S. H. (1987), Gene 61, 103–112.
Raingeaud, J., Lavergne, F., Lelievre, V., Muller, J. M., Julien, R., and Cenatiempo, Y. (1996), Biochimie 78, 14–25.
Harder, J., Bartels, J., Christophers, E., and Schröder, J. M. (1997), Nature 387, 861.
Schröder, J. M. and Harder, J. (1999), Int. J. Biochem. Cell Biol. 31, 645–651.
Ganz, T. (1999), Science 286, 420–421.
Fang, X. M., Peng, L., Xu, Z. N., Wu, J. M., and Cen, P. L. (2002), Protein Pept. Lett. 9, 31–37.
Zhang, Z. Q., Yao, L. H., and Hou, Y. D. (1990), Chin. J. Virol. 6, 111–116.
Wang, F., Fang, X. M., Xu, Z. N., Peng, L., and Cen, P. L. (2004), Prep. Biochem. Biotechnol. 34, 215–225.
Sivakesava, S., Xu, Z. N., Chen, Y. H., Hackett, J., Huang, R. C., Lam, E., Lam, T. L., Siu, K. L., Wong, R. S. C., and Wong, W. K. R. (1999), Process Biochem. 34, 893–900.
Zhao, F. Y., Bai, Y. L., Zhang, X. M., and Gao, C. C. (2001), Microbiology 28, 50–55.
Zhang, Y. K., Li, T. M., and Liu, J. J. (2003), Protein Exp. Purif. 29, 132–139.
Chao, Y. P., Chiang, C. J., and Hung, W. B. (2002), Biotechnol. Prog. 18, 394–400.
Xu, Z. N., Liu, G., Cen, P. L., and Wong, W. K. R. (2000), Bioprocess Eng. 23, 669–674.
Spehr, V., Frahm, D., and Meyer, T. F. (2000), Gene 257, 259–267.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Xu, Z., Wang, F., Peng, L. et al. Expression of human β-defensin-2 with multiple joined genes in Escherichia coli . Appl Biochem Biotechnol 120, 1–13 (2005). https://doi.org/10.1385/ABAB:120:1:01
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1385/ABAB:120:1:01