A novel PCR-based method for high throughput prokaryotic expression of antimicrobial peptide genes

BMC Biotechnology

Volumn 12, Issue , 2012, Pages

  Ke, Tao ^a,b   Liang, Su ^c   Huang, Jin ^c   Mao, Han ^b   Chen, Jibao ^a   Dong, Caihua ^b   Huang, Junyan ^b   Liu, Shengyi ^b   Kang, Jianxiong ^d   Liu, Dongqi ^d   Ma, Xiangdong ^c  

^a NANYANG NORMAL UNIVERSITY   (China)

^b INSTITUTE OF PLANT PROTECTION   (China)

^c HUBEI UNIVERSITY   (China)

^d HUAZHONG UNIVERSITY OF SCIENCE AND TECHNOLOGY   (China)

Author keywords

Antimicrobial peptide; High throughput; Npro; Prokaryotic expression

Indexed keywords

ANTI-MICROBIAL ACTIVITY; ANTIMICROBIAL PEPTIDE; BIOINFORMATIC ANALYSIS; CELL PROTEIN CONTENT; COST-EFFECTIVE METHODS; DIFFERENT SIZES; E. COLI; EXPRESSION LEVELS; EXPRESSION SYSTEM; FEVER VIRUS; FUSION PARTNERS; FUSION PROTEINS; FUSION TAG; HIGH LEVEL EXPRESSION; HIGH THROUGHPUT; HISTATIN; IN-VITRO; IN-VIVO; INCLUSION BODIES; MICROCOCCUS LUTEUS; N-TERMINALS; NPRO; OVER-EXPRESSION; PROKARYOTIC EXPRESSION; REFOLDING; SELF-CLEAVAGE; SEQUENCE DATA; SINGLE-STEP; STRUCTURAL CHARACTERIZATION; SYNTHETIC GENES;

BIOINFORMATICS; CHROMATOGRAPHY; CLONING; CYTOLOGY; ESCHERICHIA COLI; PEPTIDES; POLYPEPTIDES; PURIFICATION; THROUGHPUT; VIRUSES;

GENE EXPRESSION;

BOMBININ LIKE PEPTIDE 7; COMBI 1 PEPTIDE; HEXAPEPTIDE; HISTATIN; HISTATIN 6; HISTATIN 9; HYBRID PROTEIN; N TERMINAL AUTOPROTEASE; POLYPEPTIDE ANTIBIOTIC AGENT; PROTEIN EDDIE; TEMPORIN G; UNCLASSIFIED DRUG; ANTIMICROBIAL CATIONIC PEPTIDE; N(PRO) PROTEIN, SWINE FEVER VIRUS; PROTEINASE; VIRUS PROTEIN;

AMINO TERMINAL SEQUENCE; ANTIMICROBIAL ACTIVITY; ARTICLE; COST EFFECTIVENESS ANALYSIS; CYTOPLASM; ESCHERICHIA COLI; GENE EXPRESSION; GENE EXPRESSION SYSTEM; GENETIC RECOMBINATION; HIGH THROUGHPUT SCREENING; INCLUSION BODY MYOSITIS; MICROCOCCUS LUTEUS; MOLECULAR CLONING; NONHUMAN; POLYMERASE CHAIN REACTION; PROTEIN CLEAVAGE; PROTEIN CONTENT; PROTEIN EXPRESSION; PROTEIN PURIFICATION; PROTEIN REFOLDING; SACCHAROMYCES CEREVISIAE; BIOSYNTHESIS; CHEMISTRY; DRUG EFFECT; GENETICS; HUMAN; METABOLISM; METHODOLOGY; MICROBIOLOGICAL EXAMINATION;

CLASSICAL SWINE FEVER VIRUS; ESCHERICHIA COLI; MICROCOCCUS LUTEUS; PROKARYOTA;

ANTIMICROBIAL CATIONIC PEPTIDES; CLONING, MOLECULAR; ENDOPEPTIDASES; ESCHERICHIA COLI; HIGH-THROUGHPUT SCREENING ASSAYS; HUMANS; MICROBIAL SENSITIVITY TESTS; MICROCOCCUS LUTEUS; POLYMERASE CHAIN REACTION; PROTEIN REFOLDING; RECOMBINANT FUSION PROTEINS; SACCHAROMYCES CEREVISIAE; VIRAL PROTEINS;

EID: 84862788628     PISSN: None     EISSN: 14726750     Source Type: Journal    
DOI: 10.1186/1472-6750-12-10     Document Type: Article

References (29)

1. Koczulla AR, Bals R. Antimicrobial peptides: current status and therapeutic potential. Drugs 2003, 63(4):389-406. 10.2165/00003495-200363040-00005, 12558461.
2. Reddy KV, Yedery RD, Aranha C. Antimicrobial peptides: premises and promises. Int J Antimicrob Agents 2004, 24(6):536-547. 10.1016/j.ijantimicag.2004.09.005, 15555874.
3. Dubin A, Mak P, Dubin G, Rzychon M, Stec-Niemczyk J, Wladyka B, Maziarka K, Chmiel D. New generation of peptide antibiotics. Acta Biochim Pol 2005, 52(3):633-638.
4. Boman HG. Antibacterial peptides: key components needed in immunity. Cell 1991, 65(2):205-207. 10.1016/0092-8674(91)90154-Q, 2015623.
5. Boman HG. Peptide antibiotics and their role in innate immunity. Annu Rev Immunol 1995, 13:61-92. 10.1146/annurev.iy.13.040195.000425, 7612236.
6. Li Y. Carrier proteins for fusion expression of antimicrobial peptides in Escherichia coli. Biotechnol Appl Biochem 2009, 54(1):1-9. 10.1042/BA20090087, 19575694.
7. van't Hof W, Veerman EC, Helmerhorst EJ, Amerongen AV. Antimicrobial peptides: properties and applicability. Biol Chem 2001, 382(4):597-619.
8. Silverstein KA, Graham MA, Paape TD, VandenBosch KA. Genome organization of more than 300 defensin-like genes in Arabidopsis. Plant Physiol 2005, 138(2):600-610. 10.1104/pp.105.060079, 1150381, 15955924.
9. Pestana-Calsa MC, Ribeiro IL, Calsa T. Bioinformatics-coupled molecular approaches for unravelling potential antimicrobial peptides coding genes in Brazilian native and crop plant species. Curr Protein Pept Sci 2010, 11(3):199-209. 10.2174/138920310791112138, 20088767.
10. Xiao Y, Hughes AL, Ando J, Matsuda Y, Cheng JF, Skinner-Noble D, Zhang G. A genome-wide screen identifies a single beta-defensin gene cluster in the chicken: implications for the origin and evolution of mammalian defensins. BMC Genomics 2004, 5(1):56. 10.1186/1471-2164-5-56, 515299, 15310403.
11. Gard AL, Lenz PH, Shaw JR, Christie AE. Identification of putative peptide paracrines/hormones in the water flea Daphnia pulex (Crustacea; Branchiopoda; Cladocera) using transcriptomics and immunohistochemistry. Gen Comp Endocrinol 2009, 160(3):271-287. 10.1016/j.ygcen.2008.12.014, 19135444.
12. Christie AE. Neuropeptide discovery in Ixodoidea: an in silico investigation using publicly accessible expressed sequence tags. Gen Comp Endocrinol 2008, 157(2):174-185. 10.1016/j.ygcen.2008.03.027, 18495123.
13. Belarmino LC, Capriles PV, Crovella S, Dardene LE, Benko-Iseppon AM. EST- database search of plant defensins - an example using sugarcane, a large and complex genome. Curr Protein Pept Sci 2010, 11(3):248-254. 10.2174/138920310791112048, 20088773.
14. Silverstein KA, Moskal WA, Wu HC, Underwood BA, Graham MA, Town CD, VandenBosch KA. Small cysteine-rich peptides resembling antimicrobial peptides have been under-predicted in plants. Plant J 2007, 51(2):262-280. 10.1111/j.1365-313X.2007.03136.x, 17565583.
15. Oey M, Lohse M, Scharff LB, Kreikemeyer B, Bock R. Plastid production of protein antibiotics against pneumonia via a new strategy for high-level expression of antimicrobial proteins. Proc Natl Acad Sci USA 2009, 106(16):6579-6584. 10.1073/pnas.0813146106, 2662961, 19332784.
16. Achmuller C, Kaar W, Ahrer K, Wechner P, Hahn R, Werther F, Schmidinger H, Cserjan-Puschmann M, Clementschitsch F, Striedner G, Bayer K, Jungbauer A, Auer B. N(pro) fusion technology to produce proteins with authentic N termini in E. coli. Nat Methods 2007, 4(12):1037-1043. 10.1038/nmeth1116, 18026112.
17. Lee JH, Kim JH, Hwang SW, Lee WJ, Yoon HK, Lee HS, Hong SS. High-level expression of antimicrobial peptide mediated by a fusion partner reinforcing formation of inclusion bodies. Biochem Biophys Res Commun 2000, 277(3):575-580. 10.1006/bbrc.2000.3712, 11061996.
18. Arnau J, Lauritzen C, Petersen GE, Pedersen J. Current strategies for the use of affinity tags and tag removal for the purification of recombinant proteins. Protein Expr Purif 2006, 48(1):1-13. 10.1016/j.pep.2005.12.002, 16427311.
19. Stark R, Meyers G, Rumenapf T, Thiel HJ. Processing of pestivirus polyprotein: cleavage site between autoprotease and nucleocapsid protein of classical swine fever virus. J Virol 1993, 67(12):7088-7095. 238170, 8230432.
20. Cheng X, Lu W, Zhang S, Cao P. Expression and purification of antimicrobial peptide CM4 by Npro fusion technology in E. coli. Amino Acids 2010, 39(5):1545-1552. 10.1007/s00726-010-0625-0, 20512388.
21. Jungbauer A, Kaar W. Current status of technical protein refolding. J Biotechnol 2007, 128(3):587-596. 10.1016/j.jbiotec.2006.12.004, 17222934.
22. Zhang Z, Ke T, Zhou Y, Ma X, Ma L. High expression of antimicrobial peptide Cecropin AD in Escherichia coli by fusion with EDDIE. Sheng Wu Gong Cheng Xue Bao 2009, 25(8):1247-1253.
23. Heyman JA, Cornthwaite J, Foncerrada L, Gilmore JR, Gontang E, Hartman KJ, Hernandez CL, Hood R, Hull HM, Lee WY, Marcil R, Marsh EJ, Mudd KM, Patino MJ, Purcell TJ, Rowland JJ, Sindici ML, Hoeffler JP. Genome-scale cloning and expression of individual open reading frames using topoisomerase I-mediated ligation. Genome Res 1999, 9(4):383-392. 310734, 10207160.
24. Zhu D, Zhong X, Tan R, Chen L, Huang G, Li J, Sun X, Xu L, Chen J, Ou Y, Zhang T, Yuan D, Zhang Z, Shu W, Ma L. High-throughput cloning of human liver complete open reading frames using homologous recombination in Escherichia coli. Anal Biochem 2010, 397(2):162-167. 10.1016/j.ab.2009.10.018, 19835833.
25. Muyrers JP, Zhang Y, Stewart AF. Techniques: Recombinogenic engineering-new options for cloning and manipulating DNA. Trends Biochem Sci 2001, 26(5):325-331. 10.1016/S0968-0004(00)01757-6, 11343926.
26. Iizasa E, Nagano Y. Highly efficient yeast-based in vivo DNA cloning of multiple DNA fragments and the simultaneous construction of yeast/Escherichia coli shuttle vectors. Biotechniques 2006, 40(1):79-83. 10.2144/000112041, 16454044.
27. Olieric N, Kuchen M, Wagen S, Sauter M, Crone S, Edmondson S, Frey D, Ostermeier C, Steinmetz MO, Jaussi R. Automated seamless DNA co-transformation cloning with direct expression vectors applying positive or negative insert selection. BMC Biotechnol 2010, 10:56. 10.1186/1472-6750-10-56, 2924254, 20691119.
28. Wang Z, Wang G. APD: the Antimicrobial Peptide Database. Nucleic Acids Res 2004, 32(Database):D590-592. 308759, 14681488.
29. Lehrer RI, Rosenman M, Harwig SS, Jackson R, Eisenhauer P. Ultrasensitive assays for endogenous antimicrobial polypeptides. J Immunol Methods 1991, 137(2):167-173. 10.1016/0022-1759(91)90021-7, 1901580.