Transcriptome analyses of insect cells to facilitate baculovirus-insect expression
Received date: 31 Jan 2016
Accepted date: 27 Feb 2016
Published date: 27 May 2016
Copyright
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.
Kai Yu , Yang Yu , Xiaoyan Tang , Huimin Chen , Junyu Xiao , Xiao-Dong Su . Transcriptome analyses of insect cells to facilitate baculovirus-insect expression[J]. Protein & Cell, 2016 , 7(5) : 373 -382 . DOI: 10.1007/s13238-016-0260-y
| 1 |
Bolger AM, Lohse M, Usadel B (2014) Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 30:2114–2120.
|
| 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.
|
| 3 |
Brondyk WH (2009) Selecting an appropriate method for expressing a recombinant protein. Methods Enzymol 463:131–147.
|
| 4 |
Camacho C, Coulouris G, Avagyan V
|
| 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.
|
| 8 |
Conesa A, Götz S (2008) Blast2GO: A comprehensive suite for functional analysis in plant genomics. Int J Plant Genomics 2008:619832.
|
| 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
|
| 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
|
| 12 |
Grabherr MG, Haas BJ, Yassour M
|
| 13 |
Haas BJ, Papanicolaou A, Yassour M
|
| 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.
|
| 15 |
Hollister J, Grabenhorst E, Nimtz M
|
| 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
|
| 17 |
Holm L (1986) Codon usage and gene expression. Nucleic Acids Res 14:3075–3087
|
| 18 |
Jarvis DL (2003) Developing baculovirus-insect cell expression systems for humanized recombinant glycoprotein production. Virology 310:1–7.
|
| 19 |
Kakumani PK, Malhotra P, Mukherjee SK, Bhatnagar RK (2014) A draft genome assembly of the army worm, Spodoptera frugiperda. Genomics 104:134–143.
|
| 20 |
Kakumani PK, Shukla R, Todur VN
|
| 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.
|
| 22 |
Lairson LL, Henrissat B, Davies GJ, Withers SG (2008) Glycosyltransferases: structures, functions, and mechanisms. Annu Rev Biochem 77:521–555.
|
| 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.
|
| 24 |
Li B, Dewey CN (2011) RSEM: accurate transcript quantification from RNA-Seq data with or without a reference genome. BMC Bioinformatics 12:323.
|
| 25 |
Li H, Durbin R (2010) Fast and accurate long-read alignment with Burrows-Wheeler transform. Bioinformatics 26:589–595.
|
| 26 |
Lombard V, Golaconda Ramulu H, Drula E
|
| 27 |
Narimatsu H (2004) Construction of a human glycogene library and comprehensive functional analysis. Glycoconj J 21:17–24.
|
| 28 |
Olczak M, Olczak T (2006) Comparison of different signal peptides for protein secretion in nonlytic insect cell system. Anal Biochem 359:45–53.
|
| 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.
|
| 30 |
Powell S, Forslund K, Szklarczyk D
|
| 31 |
Soejima Y, Lee J, Nagata Y
|
| 32 |
UniProt Consortium (2015) UniProt: a hub for protein information. Nucleic Acids Res 43:D204–D212.
|
| 33 |
Vandenborre G, Smagghe G, Ghesquière B
|
| 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
|
| 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
|
| 36 |
Wickham TJ, Davis T, Granados RR,
|
| 37 |
Xie C, Mao X, Huang J
|
| 38 |
Xu C, Ng DTW(2015) Glycosylation-directed quality control of protein folding. Nat Rev Mol Cell Biol 16:742–752.
|
| 39 |
Ye J, Fang L, Zheng H
|
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| 〈 |
|
〉 |