An Overview and History of Glyco-Engineering in Insect Expression Systems

  • Christoph Geisler
  • Hideaki Mabashi-Asazuma
  • Donald L. Jarvis
Part of the Methods in Molecular Biology book series (MIMB, volume 1321)


Insect systems, including the baculovirus-insect cell and Drosophila S2 cell systems are widely used as recombinant protein production platforms. Historically, however, no insect-based system has been able to produce glycoproteins with human-type glycans, which often influence the clinical efficacy of therapeutic glycoproteins and the overall structures and functions of other recombinant glycoprotein products. In addition, some insect cell systems produce N-glycans with immunogenic epitopes. Over the past 20 years, these problems have been addressed by efforts to glyco-engineer insect-based expression systems. These efforts have focused on introducing the capacity to produce complex-type, terminally sialylated N-glycans and eliminating the capacity to produce immunogenic N-glycans. Various glyco-engineering approaches have included genetically engineering insect cells, baculoviral vectors, and/or insects with heterologous genes encoding the enzymes required to produce various glycosyltransferases, sugars, nucleotide sugars, and nucleotide sugar transporters, as well as an enzyme that can deplete GDP-fucose. In this chapter, we present an overview and history of glyco-engineering in insect expression systems as a prelude to subsequent chapters, which will highlight various methods used for this purpose.

Key words

Insect cells Baculovirus Baculovirus insect cell system Drosophila expression system Glyco-engineering Glycosylation 



Research on insect protein glycosylation pathways, the baculovirus-insect cell system, and insect expression system engineering in the authors’ labs at the University of Wyoming and GlycoBac is currently supported by National Institute of General Medical Sciences grants R44GM093411, R43GM102982, R43GM109504, and National Institute of Allergy and Infectious Diseases grant R43AI112118. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institute of General Medical Sciences, the National Institute of Allergy and Infectious Disease, or the National Institutes of Health.


  1. 1.
    Smith GE, Summers MD, Fraser MJ (1983) Production of human beta interferon in insect cells infected with a baculovirus expression vector. Mol Cell Biol 3:2156–2165PubMedCentralPubMedGoogle Scholar
  2. 2.
    Pennock GD, Shoemaker C, Miller LK (1984) Strong and regulated expression of Escherichia coli beta-galactosidase in insect cells with a baculovirus vector. Mol Cell Biol 4:399–406PubMedCentralPubMedGoogle Scholar
  3. 3.
    Rio DC, Rubin GM (1985) Transformation of cultured Drosophila melanogaster cells with a dominant selectable marker. Mol Cell Biol 5:1833–1838PubMedCentralPubMedGoogle Scholar
  4. 4.
    Johansen H, van der Straten A, Sweet R et al (1989) Regulated expression at high copy number allows production of a growth-inhibitory oncogene product in Drosophila Schneider cells. Gene Dev 3:882–889PubMedCrossRefGoogle Scholar
  5. 5.
    Jarvis DL (1997) Baculovirus expression vectors. In: Miller LK (ed) The baculoviruses. Plenum, New York, pp 389–431CrossRefGoogle Scholar
  6. 6.
    Jarvis DL (2009) Baculovirus-insect cell expression systems. Methods Enzymol 463:191–222PubMedGoogle Scholar
  7. 7.
    Maeda S (1989) Expression of foreign genes in insects using baculovirus vectors. Annu Rev Entomol 34:351–372PubMedCrossRefGoogle Scholar
  8. 8.
    Usami A, Suzuki T, Nagaya H et al (2010) Silkworm as a host of baculovirus expression. Curr Pharm Biotechnol 11:246–250PubMedCrossRefGoogle Scholar
  9. 9.
    Choudary PV, Kamita SG, Maeda S (1995) Expression of foreign genes in Bombyx mori larvae using baculovirus vectors. Methods Mol Biol 39:243–264PubMedGoogle Scholar
  10. 10.
    Kato T, Kajikawa M, Maenaka K et al (2010) Silkworm expression system as a platform technology in life science. Appl Microbiol Biotechnol 85:459–470PubMedCentralPubMedCrossRefGoogle Scholar
  11. 11.
    Lynn DE (2007) Available lepidopteran insect cell lines. Methods Mol Biol 388:117–138PubMedGoogle Scholar
  12. 12.
    Jarvis DL (1993) Continuous foreign gene expression in stably-transformed insect cells. In: Goosen MFA, Daugulis A, Faulkner P (eds) Insect cell culture engineering. Marcel Dekker Inc, New York, pp 193–217Google Scholar
  13. 13.
    Schetz JA, Shankar EP (2004) Protein expression in the Drosophila Schneider 2 cell system. Curr Protoc Neurosci. Chapter 4, Unit 4, 16Google Scholar
  14. 14.
    Douris V, Swevers L, Labropoulou V et al (2006) Stably transformed insect cell lines: tools for expression of secreted and membrane-anchored proteins and high-throughput screening platforms for drug and insecticide discovery. Adv Virus Res 68:113–156PubMedGoogle Scholar
  15. 15.
    Cherbas L, Cherbas P (2007) Transformation of Drosophila cell lines: an alternative approach to exogenous protein expression. Methods Mol Biol 388:317–340PubMedGoogle Scholar
  16. 16.
    Pfeifer TA (1998) Expression of heterologous proteins in stable insect cell culture. Curr Opin Biotechnol 9:518–521PubMedCrossRefGoogle Scholar
  17. 17.
    Katoh T, Tiemeyer M (2013) The N’s and O’s of Drosophila glycoprotein glycobiology. Glycoconj J 30:57–66PubMedCentralPubMedCrossRefGoogle Scholar
  18. 18.
    ten Hagen KG, Zhang L, Tian E et al (2009) Glycobiology on the fly: developmental and mechanistic insights from Drosophila. Glycobiology 19:102–111PubMedCentralPubMedCrossRefGoogle Scholar
  19. 19.
    Marz L, Altmann F, Staudacher E et al (1995) Protein glycosylation in insects. In: Montreuil J, Vliegenthart JFG, Schachter H (eds) Glycoproteins, vol 29a. Elsevier, Amsterdam, pp 543–563CrossRefGoogle Scholar
  20. 20.
    Altmann F, Staudacher E, Wilson IB et al (1999) Insect cells as hosts for the expression of recombinant glycoproteins. Glycoconj J 16:109–123PubMedCrossRefGoogle Scholar
  21. 21.
    Marchal I, Jarvis DL, Cacan R et al (2001) Glycoproteins from insect cells: sialylated or not? Biol Chem 382:151–159PubMedCentralPubMedCrossRefGoogle Scholar
  22. 22.
    Tomiya N, Narang S, Lee YC et al (2004) Comparing N-glycan processing in mammalian cell lines to native and engineered lepidopteran insect cell lines. Glycoconj J 21:343–360PubMedCrossRefGoogle Scholar
  23. 23.
    Harrison RL, Jarvis DL (2006) Protein N-glycosylation in the baculovirus-insect cell expression system and engineering of insect cells to produce “mammalianized” recombinant glycoproteins. Adv Virus Res 68:159–191PubMedGoogle Scholar
  24. 24.
    Kim K, Lawrence SM, Park J et al (2002) Expression of a functional Drosophila melanogaster N-acetylneuraminic acid (Neu5Ac) phosphate synthase gene: evidence for endogenous sialic acid biosynthetic ability in insects. Glycobiology 12:73–83PubMedCrossRefGoogle Scholar
  25. 25.
    Koles K, Irvine KD, Panin VM (2004) Functional characterization of a Drosophila sialyltransferase. J Biol Chem 279:4346–4357PubMedCrossRefGoogle Scholar
  26. 26.
    Viswanathan K, Tomiya N, Singh S et al (2006) Expression of a functional Drosophila melanogaster CMP-sialic acid synthetase: differential localization of the Drosophila and human enzymes. J Biol Chem 281:15929–15940PubMedCrossRefGoogle Scholar
  27. 27.
    Aoki K, Perlman M, Lim JM et al (2007) Dynamic developmental elaboration of N-linked glycan complexity in the Drosophila melanogaster embryo. J Biol Chem 282:9127–9142PubMedCrossRefGoogle Scholar
  28. 28.
    Koles K, Lim JM, Aoki K et al (2007) Identification of N-glycosylated proteins from the central nervous system of Drosophila melanogaster. Glycobiology 17:1388–1403PubMedCrossRefGoogle Scholar
  29. 29.
    Repnikova E, Koles K, Nakamura M et al (2010) Sialyltransferase regulates nervous system function in Drosophila. J Neurosci 30:6466–6476PubMedCentralPubMedCrossRefGoogle Scholar
  30. 30.
    Altmann F, Schwihla H, Staudacher E et al (1995) Insect cells contain an unusual, membrane-bound ß-N-acetylglucosaminidase probably involved in the processing of protein N-glycans. J Biol Chem 270:17344–17349PubMedCrossRefGoogle Scholar
  31. 31.
    Leonard R, Rendic D, Rabouille C et al (2006) The Drosophila fused lobes gene encodes an N-acetylglucosaminidase involved in N-glycan processing. J Biol Chem 281:4867–4875PubMedCrossRefGoogle Scholar
  32. 32.
    Geisler C, Aumiller JJ, Jarvis DL (2008) A fused lobes gene encodes the processing ß-N-acetylglucosaminidase in Sf9 cells. J Biol Chem 283:11330–11339PubMedCentralPubMedCrossRefGoogle Scholar
  33. 33.
    Geisler C, Jarvis DL (2010) Identification of genes encoding N-glycan processing ß-N-acetylglucosaminidases in Trichoplusia ni and Bombyx mori: implications for glyco-engineering of baculovirus expression systems. Biotechnol Prog 26:34–44PubMedCentralPubMedGoogle Scholar
  34. 34.
    Kulakosky PC, Hughes PR, Wood HA (1998) N-linked glycosylation of a baculovirus-expressed recombinant glycoprotein in insect larvae and tissue culture cells. Glycobiology 8:741–745PubMedCrossRefGoogle Scholar
  35. 35.
    Park EY, Ishikiriyama M, Nishina T et al (2009) Human IgG1 expression in silkworm larval hemolymph using BmNPV bacmids and its N-linked glycan structure. J Biotechnol 139:108–114PubMedCrossRefGoogle Scholar
  36. 36.
    Sasaki K, Kajikawa M, Kuroki K et al (2009) Silkworm expression and sugar profiling of human immune cell surface receptor, KIR2DL1. Biochem Biophys Res Commun 387:575–580PubMedCrossRefGoogle Scholar
  37. 37.
    Lin SC, Jan JT, Dionne B et al (2013) Different immunity elicited by recombinant H5N1 hemagglutinin proteins containing pauci-mannose, high-mannose, or complex type N-glycans. PLoS One 8:e66719PubMedCentralPubMedCrossRefGoogle Scholar
  38. 38.
    Watanabe S, Kokuho T, Takahashi H et al (2001) Sialylation of N-glycans on the recombinant proteins expressed by a baculovirus-insect cell system under ß-N-acetylglucosaminidase inhibition. J Biol Chem 277:5090–5093PubMedCrossRefGoogle Scholar
  39. 39.
    Kim YK, Kim KR, Kang DG et al (2009) Suppression of ß-N-acetylglucosaminidase in the N-glycosylation pathway for complex glycoprotein formation in Drosophila S2 cells. Glycobiology 19:301–308PubMedCrossRefGoogle Scholar
  40. 40.
    Kim YK, Kim KR, Kang DG et al (2011) Expression of ß-1,4-galactosyltransferase and suppression of ß-N-acetylglucosaminidase to aid synthesis of complex N-glycans in insect Drosophila S2 cells. J Biotech 153:145–152CrossRefGoogle Scholar
  41. 41.
    Chang KH, Yang JM, Chun HO et al (2005) Enhanced activity of recombinant ß-secretase from Drosophila melanogaster S2 cells transformed with cDNAs encoding human ß1,4-galactosyltransferase and Galß1,4-GlcNAc α2,6-sialyltransferase. J Biotechnol 116:359–367PubMedCrossRefGoogle Scholar
  42. 42.
    Hu JB, Zhang P, Wang MX et al (2012) A transgenic Bm cell line of piggyBac transposon-derived targeting expression of humanized glycoproteins through N-glycosylation. Mol Biol Rep 39:8405–8413PubMedCrossRefGoogle Scholar
  43. 43.
    Kidd IM, Emery VC (1993) The use of baculoviruses as expression vectors. Appl Biochem Biotechnol 42:137–159PubMedCrossRefGoogle Scholar
  44. 44.
    Roy P (1996) Genetically engineered particulate virus-like structures and their use as vaccine delivery systems. Intervirology 39:62–71PubMedGoogle Scholar
  45. 45.
    Fitzgerald DJ, Berger P, Schaffitzel C et al (2006) Protein complex expression by using multigene baculoviral vectors. Nat Methods 3:1021–1032PubMedCrossRefGoogle Scholar
  46. 46.
    Harrison RL, Jarvis DL (2007) Transforming lepidopteran insect cells for continuous recombinant protein expression. Methods Mol Biol 388:299–316PubMedGoogle Scholar
  47. 47.
    Harrison RL, Jarvis DL (2007) Transforming lepidopteran insect cells for improved protein processing. Methods Mol Biol 388:341–356PubMedGoogle Scholar
  48. 48.
    Toth AM, Kuo CW, Khoo KH et al (2014) A new insect cell glyco-engineering approach provides baculovirus-inducible glycogene expression and increases human-type glycosylation efficiency. J Biotechnol 182–183:19–29PubMedCrossRefGoogle Scholar
  49. 49.
    Hollister JR, Shaper JH, Jarvis DL (1998) Stable expression of mammalian ß1,4-galactosyltransferase extends the N-glycosylation pathway in insect cells. Glycobiology 8:473–480PubMedCrossRefGoogle Scholar
  50. 50.
    Breitbach K, Jarvis DL (2001) Improved glycosylation of a foreign protein by Tn-5B1-4 cells engineered to express mammalian glycosyltransferases. Biotechnol Bioeng 74:230–239PubMedCentralPubMedCrossRefGoogle Scholar
  51. 51.
    Jarvis DL, Fleming JA, Kovacs GR et al (1990) Use of early baculovirus promoters for continuous expression and efficient processing of foreign gene products in stably transformed lepidopteran cells. Nat Biotechnol 8:950–955CrossRefGoogle Scholar
  52. 52.
    Culp JS, Johansen H, Hellmig B et al (1991) Regulated expression allows high level production and secretion of HIV-1 gp120 envelope glycoprotein in Drosophila Schneider cells. Nat Biotechnol 9:173–177CrossRefGoogle Scholar
  53. 53.
    Dorer DR, Henikoff S (1997) Transgene repeat arrays interact with distant heterochromatin and cause silencing in cis and trans. Genetics 147:1181–1190PubMedCentralPubMedGoogle Scholar
  54. 54.
    Pfeifer TA, Hegedus DD, Grigliatti TA et al (1997) Baculovirus immediate-early promoter-mediated expression of the Zeocin resistance gene for use as a dominant selectable marker in dipteran and lepidopteran insect cell lines. Gene 188:183–190PubMedCrossRefGoogle Scholar
  55. 55.
    Farrell PJ, Lu M, Prevost J, Brown C et al (1998) High-level expression of secreted glycoproteins in transformed lepidopteran insect cells using a novel expression vector. Biotechnol Bioeng 60:656–663PubMedCrossRefGoogle Scholar
  56. 56.
    Aumiller JJ, Mabashi-Asazuma H, Hillar A et al (2012) A new glyco-engineered insect cell line with an inducibly mammalianized protein N-glycosylation pathway. Glycobiology 22:417–428PubMedCentralPubMedCrossRefGoogle Scholar
  57. 57.
    Mabashi-Asazuma H, Kuo CW, Khoo KH et al (2014) A novel baculovirus vector for the production of nonfucosylated recombinant glycoproteins in insect cells. Glycobiology 24:325–340PubMedCentralPubMedCrossRefGoogle Scholar
  58. 58.
    Wagner R, Liedtke S, Kretzschmar E et al (1996) Elongation of the N-glycans of fowl plague virus hemagglutinin expressed in Spodoptera frugiperda (Sf9) cells by coexpression of human ß1,2-N-acetylglucosaminyltransferase I. Glycobiology 6:165–175Google Scholar
  59. 59.
    Jarvis DL, Finn EE (1996) Modifying the insect cell N-glycosylation pathway with immediate early baculovirus expression vectors. Nat Biotechnol 14:1288–1292PubMedCrossRefGoogle Scholar
  60. 60.
    Wolff MW, Murhammer DW, Jarvis DL et al (1999) Electrophoretic analysis of glycoprotein glycans produced by lepidopteran insect cells infected with an immediate early recombinant baculovirus encoding mammalian ß1,4-galactosyltransferase. Glycoconj J 16:753–756PubMedCrossRefGoogle Scholar
  61. 61.
    Ailor E, Takahashi N, Tsukamoto Y et al (2000) N-glycan patterns of human transferrin produced in Trichoplusia ni insect cells: effects of mammalian galactosyltransferase. Glycobiology 10:837–847PubMedCrossRefGoogle Scholar
  62. 62.
    Palmberger D, Wilson IB, Berger I et al (2012) SweetBac: a new approach for the production of mammalianised glycoproteins in insect cells. PLoS One 7:e34226PubMedCentralPubMedCrossRefGoogle Scholar
  63. 63.
    Tomiya N, Howe D, Aumiller JJ et al (2003) Complex-type biantennary N-glycans of recombinant human transferrin from Trichoplusia ni insect cells expressing mammalian ß1,4-galactosyltransferase and ß1, 2-N-acetylglucosaminyltransferase II. Glycobiology 13:23–34Google Scholar
  64. 64.
    Seo NS, Hollister JR, Jarvis DL (2001) Mammalian glycosyltransferase expression allows sialoglycoprotein production by baculovirus-infected insect cells. Protein Expr Purif 22:234–241PubMedCrossRefGoogle Scholar
  65. 65.
    Hooker AD, Green NH, Baines AJ et al (1999) Constraints on the transport and glycosylation of recombinant IFN-γ in Chinese hamster ovary and insect cells. Biotechnol Bioeng 63:559–572PubMedCrossRefGoogle Scholar
  66. 66.
    Tomiya N, Ailor E, Lawrence SM et al (2001) Determination of nucleotides and sugar nucleotides involved in protein glycosylation by high-performance anion-exchange chromatography: sugar nucleotide contents in cultured insect cells and mammalian cells. Anal Biochem 293:129–137PubMedCrossRefGoogle Scholar
  67. 67.
    Hollister JR, Conradt HO, Jarvis DL (2003) Evidence for a sialic acid salvaging pathway in lepidopteran insect cell lines. Glycobiology 13:487–495PubMedCentralPubMedCrossRefGoogle Scholar
  68. 68.
    Jarvis DL, Howe D, Aumiller JJ (2001) Novel baculovirus expression vectors that provide sialylation of recombinant glycoproteins in lepidopteran insect cells. J Virol 75:6223–6227PubMedCentralPubMedCrossRefGoogle Scholar
  69. 69.
    Chang GD, Chen CJ, Lin CY et al (2003) Improvement of glycosylation in insect cells with mammalian glycosyltransferases. J Biotechnol 102:61–71CrossRefGoogle Scholar
  70. 70.
    Hill DR, Aumiller JJ, Shi X et al (2006) Isolation and analysis of a baculovirus vector that supports recombinant glycoprotein sialylation by SfSWT-1 cells cultured in serum-free medium. Biotechnol Bioeng 95:37–47PubMedCentralPubMedCrossRefGoogle Scholar
  71. 71.
    Hollister JR, Grabenhorst E, Nimtz M et al (2002) Engineering the protein N-glycosylation pathway in insect cells for production of biantennary, complex N-glycans. Biochemistry 41:15093–15104PubMedCentralPubMedCrossRefGoogle Scholar
  72. 72.
    von Horsten HH, Ogorek C, Blanchard V et al (2010) Production of non-fucosylated antibodies by co-expression of heterologous GDP-6-deoxy-D-lyxo-4-hexulose reductase. Glycobiology 20:1607–1618CrossRefGoogle Scholar
  73. 73.
    Shields RL, Lai J, Keck R et al (2002) Lack of fucose on human IgG1 N-linked oligosaccharide improves binding to human Fcγ RIII and antibody-dependent cellular toxicity. J Biol Chem 277:26733–26740PubMedCrossRefGoogle Scholar
  74. 74.
    Palmberger D, Ashjaei K, Strell S et al (2014) Minimizing fucosylation in insect cell-derived glycoproteins reduces binding to IgE antibodies from the sera of patients with allergy. Biotechnol J 9:1206–1214. doi: 10.1002/biot.201300061 PubMedCrossRefGoogle Scholar
  75. 75.
    Blissard GW, Rohrmann GF (1989) Location, sequence, transcriptional mapping, and temporal expression of the gp64 envelope glycoprotein gene of the Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus. Virology 170:537–555PubMedCrossRefGoogle Scholar
  76. 76.
    Jarvis DL, Garcia A Jr (1994) Biosynthesis and processing of the Autographa californica nuclear polyhedrosis virus gp64 protein. Virology 205:300–313PubMedCrossRefGoogle Scholar
  77. 77.
    Hollister J, Jarvis DL (2001) Engineering lepidopteran insect cells for sialoglycoprotein production by genetic transformation with mammalian ß1,4-galactosyltransferase and α2,6-sialyltransferase genes. Glycobiology 11:1–9PubMedCrossRefGoogle Scholar
  78. 78.
    Yun EY, Goo TW, Kim SW et al (2005) Galactosylation and sialylation of mammalian glycoproteins produced by baculovirus-mediated gene expression in insect cells. Biotechnol Lett 27:1035–1039PubMedCrossRefGoogle Scholar
  79. 79.
    Geisler C, Jarvis DL (2012) Innovative use of a bacterial enzyme involved in sialic acid degradation to initiate sialic acid biosynthesis in glyco-engineered insect cells. Metab Eng 14:642–652PubMedCentralPubMedCrossRefGoogle Scholar
  80. 80.
    Shi X, Harrison RL, Hollister JR et al (2007) Construction and characterization of new piggyBac vectors for constitutive or inducible expression of heterologous gene pairs and the identification of a previously unrecognized activator sequence in piggyBac. BMC Biotechnol 7:5PubMedCentralPubMedCrossRefGoogle Scholar
  81. 81.
    Altmann F, Kornfeld G, Dalik T et al (1993) Processing of asparagine-linked oligosaccharides in insect cells. N-acetylglucosaminyltransferase I and II activities in cultured lepidopteran cells. Glycobiology 3:619–625PubMedCrossRefGoogle Scholar
  82. 82.
    Okada T, Ihara H, Ito R et al (2010) N-glycosylation engineering of lepidopteran insect cells by the introduction of the ß1,4-N-acetylglucosaminyltransferase III gene. Glycobiology 20:1147–1159Google Scholar
  83. 83.
    Aumiller JJ, Hollister JR, Jarvis DL (2003) A transgenic lepidopteran insect cell line engineered to produce CMP-sialic acid and sialoglycoproteins. Glycobiology 13:497–507PubMedCentralPubMedCrossRefGoogle Scholar
  84. 84.
    Munster AK, Eckhardt M, Potvin B et al (1998) Mammalian cytidine 5′-monophosphate N-acetylneuraminic acid synthetase: a nuclear protein with evolutionarily conserved structural motifs. Proc Natl Acad Sci U S A 95:9140–9145PubMedCentralPubMedCrossRefGoogle Scholar
  85. 85.
    Koles K, Repnikova E, Pavlova G et al (2009) Sialylation in protostomes: a perspective from Drosophila genetics and biochemistry. Glycoconj J 26:313–324PubMedCrossRefGoogle Scholar
  86. 86.
    Mabashi-Asazuma H, Shi X, Geisler C et al (2013) Impact of a human CMP-sialic acid transporter on recombinant glycoprotein sialylation in glyco-engineered insect cells. Glycobiology 23:199–210PubMedCentralPubMedCrossRefGoogle Scholar
  87. 87.
    Lin CH, Jarvis DL (2013) Utility of temporally distinct baculovirus promoters for constitutive and baculovirus-inducible transgene expression in transformed insect cells. J Biotechnol 165:11–17PubMedCentralPubMedCrossRefGoogle Scholar
  88. 88.
    Kim NY, Baek JY, Choi HS et al (2012) Short-hairpin RNA-mediated gene expression interference in Trichoplusia ni cells. J Microbiol Biotechnol 22:190–198PubMedCrossRefGoogle Scholar
  89. 89.
    Tamura T, Thibert C, Royer C et al (2000) Germline transformation of the silkworm Bombyx mori L. using a piggyBac transposon-derived vector. Nat Biotechnol 18:81–84PubMedCrossRefGoogle Scholar
  90. 90.
    Tomita M (2011) Transgenic silkworms that weave recombinant proteins into silk cocoons. Biotechnol Lett 3:645–654CrossRefGoogle Scholar
  91. 91.
    Fraser MJ Jr (2012) Insect transgenesis: current applications and future prospects. Annu Rev Entomol 57:267–289PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Christoph Geisler
    • 1
  • Hideaki Mabashi-Asazuma
    • 2
  • Donald L. Jarvis
    • 1
    • 2
  1. 1.GlycoBac, LLCLaramieUSA
  2. 2.Department of Molecular BiologyUniversity of WyomingLaramieUSA

Personalised recommendations