Abstract
A cDNA coding mutated cecropin CMIV fromBombyx mori was synthesized according to its amino acid sequence usingE. coli biased codons. The gene was cloned into the fusion expression vector pEZZ318 and was expressed inE. coli HB101. The fusion protein produced was purified by affinity chromatography to yield 26 mg/L fusion product. The anti-bacterial activities of recombinant cecropin CMIV were recovered after cleavage by chemical method.
Similar content being viewed by others
References
Zhang Shuangquan, Qu Xianming, Qi Zhengwu, Insect immune repines and the applicational prospects of cecropins,J. Biochem. (in Chinese), 1987, 3(1): 11.
Anderson, D., Engstrom, A., Josephson, S.et al., Biologically active and amidated cecropin produced in a baculovirus expression system from a fusion construct containing the antibody-binding part of protein A,Biochem. J., 1991, 280:219.
Hellers, M., Gunne, H., Steiner, H., Expression and post-translational processing of preprocecropin A using a baculovirus vector,Eur. J. Biochem., 1991, 199:435.
Ma Lijing, Zhuang Chuxiong, Huang Ziranet al., Transgenic yeast with cecropin D fromAtheraea pernyia and analysis of the expression product,Canye Kexue (in Chinese), 1995, 21:43.
Qu Xianming, Wu Kezuo, Qiu Xuezhenet al., Isolation and characterization of six antibacterial peptides from the hemolymph of immunizedBombyx mori pupae by injection of poly I: C,Ada Biochimica et Biophisica Academia (in Chinese), 1986, 18(3): 284.
Xie Wei, Qiu Qifeng, Chen Jiangninget al., Chemical synthesis and cloning of cecropin CMIV gene from Chinese silkworm,J. Nanjing University (natural science) (in Chinese), 1996, 32(3): 414.
Nilsson, B., Holmgren, E., Josephson, S.et al., Efficient secretion and purification of human insulin-like growth factor I with a gene fusion vector in Staphylococci,Nuclear Acids Res., 1985, 13:1151.
Moks, T., Abrahmsen, L., Holmgren, E.et al., Expression of human insulin-like growth factor I in bacteria: use of optimized gene fusion vectors to facilitate protein purification,Biochemistry, 1987, 26:5239.
Sun, N.E., Shen, B.H., Zhou, J.M.et al., An efficient method of large-scale isolation of plasmid DNAs by heat-alkali codenaturation,DNA Cell Biol., 1994, 13:83.
Sambrook, J., Fritsh, E.F., Maniatis, T.,Molecular Cloning; A Laboratory Manual, 2nd Ed., New York: Cold Spring Harbor Laboratory Press, 1989.
Hammarberg, B., Nygren, P.A., Holmgren, E.et al., Dual affinity fusion approach and its use to express recombinant human insulin-like growth factor I,Proc. Natl. Acad. Sci. USA, 1989, 86:4367.
Doherty, A. J., Connolly, B. A., Worral, A.F., Overproduction of the toxic protein bovine pancreatic DNase Iin E, coli using a tightly controlled T7-promoter-based vector,Gene, 1993, 136:337.
Uhlen, M., Moks, T., Gene fusion’s purpose of expression: An introduction,Methods in Enzymology, 1990, 185:129.
Hultmark, D., Engstrom, A., Bennich, H.et al., Insect immunity: isolation and structure of cecropin D and four minor antibacterial components from cecropia pupae,Eur. J. Biochem., 1982, 127:207.
Hua, Z., Li, J., Zhu, D., Expression of a fibrinolytically active human pro-urokinase fusion protein inE. coli., Biochem. and Molecular International, 1994, 33:1215.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Xie, W., Qiu, Q., Dong, X. et al. Fusion expression of mutated cecropin CMIV inE. coli . Sci. China Ser. C.-Life Sci. 40, 225–231 (1997). https://doi.org/10.1007/BF02879080
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF02879080