Transcriptome analyses of insect cells to facilitate baculovirus-insect expression
Kai Yu, Yang Yu, Xiaoyan Tang, Huimin Chen, Junyu Xiao, Xiao-Dong Su
Transcriptome analyses of insect cells to facilitate baculovirus-insect expression
The High Five cell line (BTI-TN-5B1-4) isolated from the cabbage looper, Trichoplusia ni is an insect cell line widely used for baculovirus-mediated recombinant protein expression. Despite its widespread application in industry and academic laboratories, the genomic background of this cell line remains unclear. Here we sequenced the transcriptome of High Five cells and assembled 25,234 transcripts. Codon usage analysis showed that High Five cells have a robust codon usage capacity and therefore suit for expressing proteins of both eukaryotic- and prokaryotic-origin. Genes involved in glycosylation were profiled in our study, providing guidance for engineering glycosylated proteins in the insect cells. We also predicted signal peptides for transcripts with high expression abundance in both High Five and Sf21 cell lines, and these results have important implications for optimizing the expression level of some secretory and membrane proteins.
High Five cell line / baculovirus-insect cell system / codon usage / glycosylation / signal peptide
[1] |
Bolger AM, Lohse M, Usadel B (2014) Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 30:2114–2120.
CrossRef
Google scholar
|
[2] |
Breitbach K, Jarvis DL (2001) Improved glycosylation of a foreign protein by Tn-5B1-4 cells engineered to express mammalian glycosyltransferases. Biotechnol Bioeng 74:230–239.
CrossRef
Google scholar
|
[3] |
Brondyk WH (2009) Selecting an appropriate method for expressing a recombinant protein. Methods Enzymol 463:131–147.
CrossRef
Google scholar
|
[4] |
Camacho C, Coulouris G, Avagyan V
CrossRef
Google scholar
|
[5] |
Cannarozzi GM, Schneider A (eds) (2012) Codon Evolution. Oxford University Press
|
[6] |
Castilho A (ed) (2015) Glyco-Engineering. Springer New York, New York, NY
|
[7] |
Chaney JL, Clark PL (2015) Roles for Synonymous Codon Usage in Protein Biogenesis. Annu Rev Biophys 44:143–166.
CrossRef
Google scholar
|
[8] |
Conesa A, Götz S (2008) Blast2GO: A comprehensive suite for functional analysis in plant genomics. Int J Plant Genomics 2008:619832.
CrossRef
Google scholar
|
[9] |
Davis TR, Trotter KM, Granados RR, Wood HA (1992) Baculovirus Expression of Alkaline Phosphatase as a Reporter Gene for Evaluation of Production, Glycosylation and Secretion. Bio/Technology 10:1148–1150. doi:10.1038/nbt1092-1148
CrossRef
Google scholar
|
[10] |
Finn RD, Bateman A, Clements J et al (2014) Pfam: the protein families database. Nucleic Acids Res 42:D222–D230. doi:10.1093/nar/gkt1223
|
[11] |
Fu L, Niu B, Zhu Z
CrossRef
Google scholar
|
[12] |
Grabherr MG, Haas BJ, Yassour M
CrossRef
Google scholar
|
[13] |
Haas BJ, Papanicolaou A, Yassour M
CrossRef
Google scholar
|
[14] |
Hollister JR, Jarvis DL (2001) Engineering lepidopteran insect cells for sialoglycoprotein production by genetic transformation with mammalian 1,4-galactosyltransferase and 2,6-sialyltransferase genes. Glycobiology 11:1–9.
CrossRef
Google scholar
|
[15] |
Hollister J, Grabenhorst E, Nimtz M
CrossRef
Google scholar
|
[16] |
Hollister JR, Shaper JH, Jarvis DL (1998) Stable expression of mammalian beta 1,4-galactosyltransferase extends the N-glycosylation pathway in insect cells. Glycobiology 8:473–480
CrossRef
Google scholar
|
[17] |
Holm L (1986) Codon usage and gene expression. Nucleic Acids Res 14:3075–3087
CrossRef
Google scholar
|
[18] |
Jarvis DL (2003) Developing baculovirus-insect cell expression systems for humanized recombinant glycoprotein production. Virology 310:1–7.
CrossRef
Google scholar
|
[19] |
Kakumani PK, Malhotra P, Mukherjee SK, Bhatnagar RK (2014) A draft genome assembly of the army worm, Spodoptera frugiperda. Genomics 104:134–143.
CrossRef
Google scholar
|
[20] |
Kakumani PK, Shukla R, Todur VN
CrossRef
Google scholar
|
[21] |
Kost TA, Condreay JP, Jarvis DL (2005) Baculovirus as versatile vectors for protein expression in insect and mammalian cells. Nat Biotechnol 23:567–575.
CrossRef
Google scholar
|
[22] |
Lairson LL, Henrissat B, Davies GJ, Withers SG (2008) Glycosyltransferases: structures, functions, and mechanisms. Annu Rev Biochem 77:521–555.
CrossRef
Google scholar
|
[23] |
Langmead B, Trapnell C, Pop M, Salzberg SL (2009) Ultrafast and memory-efficient alignment of short DNA sequences to the human genome. Genome Biol 10:R25.
CrossRef
Google scholar
|
[24] |
Li B, Dewey CN (2011) RSEM: accurate transcript quantification from RNA-Seq data with or without a reference genome. BMC Bioinformatics 12:323.
CrossRef
Google scholar
|
[25] |
Li H, Durbin R (2010) Fast and accurate long-read alignment with Burrows-Wheeler transform. Bioinformatics 26:589–595.
CrossRef
Google scholar
|
[26] |
Lombard V, Golaconda Ramulu H, Drula E
CrossRef
Google scholar
|
[27] |
Narimatsu H (2004) Construction of a human glycogene library and comprehensive functional analysis. Glycoconj J 21:17–24.
CrossRef
Google scholar
|
[28] |
Olczak M, Olczak T (2006) Comparison of different signal peptides for protein secretion in nonlytic insect cell system. Anal Biochem 359:45–53.
CrossRef
Google scholar
|
[29] |
Petersen TN, Brunak S, von Heijne G, Nielsen H (2011) SignalP 4.0: discriminating signal peptides from transmembrane regions. Nat Methods 8:785–786.
CrossRef
Google scholar
|
[30] |
Powell S, Forslund K, Szklarczyk D
CrossRef
Google scholar
|
[31] |
Soejima Y, Lee J, Nagata Y
CrossRef
Google scholar
|
[32] |
UniProt Consortium (2015) UniProt: a hub for protein information. Nucleic Acids Res 43:D204–D212.
CrossRef
Google scholar
|
[33] |
Vandenborre G, Smagghe G, Ghesquière B
CrossRef
Google scholar
|
[34] |
Vaughn JL, Goodwin RH, Tompkins GJ, McCawley P (1977) The Establishment of Two Cell Lines from the Insect Spodoptera frugiperda (Lepidoptera; Noctuidae). In Vitro 13:213–217
CrossRef
Google scholar
|
[35] |
von Heijne G, Abrahmsén L (1989) Species-specific variation in signal peptide design. Implications for protein secretion in foreign hosts. FEBS Lett 244:439–446
CrossRef
Google scholar
|
[36] |
Wickham TJ, Davis T, Granados RR,
CrossRef
Google scholar
|
[37] |
Xie C, Mao X, Huang J
CrossRef
Google scholar
|
[38] |
Xu C, Ng DTW(2015) Glycosylation-directed quality control of protein folding. Nat Rev Mol Cell Biol 16:742–752.
CrossRef
Google scholar
|
[39] |
Ye J, Fang L, Zheng H
CrossRef
Google scholar
|
/
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