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Fusion expression of mutated cecropin CMIV inE. coli

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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.

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References

  1. Zhang Shuangquan, Qu Xianming, Qi Zhengwu, Insect immune repines and the applicational prospects of cecropins,J. Biochem. (in Chinese), 1987, 3(1): 11.

    Google Scholar 

  2. 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.

    Google Scholar 

  3. Hellers, M., Gunne, H., Steiner, H., Expression and post-translational processing of preprocecropin A using a baculovirus vector,Eur. J. Biochem., 1991, 199:435.

    Article  PubMed  CAS  Google Scholar 

  4. 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.

    Google Scholar 

  5. 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.

    Google Scholar 

  6. 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.

    Google Scholar 

  7. 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.

    Article  CAS  Google Scholar 

  8. 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.

    Article  PubMed  CAS  Google Scholar 

  9. 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.

    PubMed  CAS  Google Scholar 

  10. Sambrook, J., Fritsh, E.F., Maniatis, T.,Molecular Cloning; A Laboratory Manual, 2nd Ed., New York: Cold Spring Harbor Laboratory Press, 1989.

    Google Scholar 

  11. 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.

    Article  PubMed  CAS  Google Scholar 

  12. 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.

    Article  PubMed  CAS  Google Scholar 

  13. Uhlen, M., Moks, T., Gene fusion’s purpose of expression: An introduction,Methods in Enzymology, 1990, 185:129.

    Article  PubMed  CAS  Google Scholar 

  14. 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.

    Article  PubMed  CAS  Google Scholar 

  15. 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.

    CAS  Google Scholar 

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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

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  • DOI: https://doi.org/10.1007/BF02879080

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