Expression of Ericerus pela Wax Ester Synthase in Spodoptera frugiperda Sf9 Cell Line

QI Qian; YU Shu-hui; SUN Tao; WANG Xue-qing; LIU Bo-wen; YANG Pu; CHEN Xiao-ming

Volume 29 Issue 2

Article Contents

Turn off MathJax

Article Navigation > Forest Research > 2016 > 29(2): 191-195

Citation:

Expression of Ericerus pela Wax Ester Synthase in Spodoptera frugiperda Sf9 Cell Line

1.
Research Institute of Resources Insects, Chinese Academy of Forestry, Key laboratory of Cultivating and Utilization of Resources Insects of State Forestry Administration, Kunming 650224, Yunnan, China

Received Date: 2015-11-15

Abstract

[Objective]Use Bac-to-Bac expression system to express WS in Sf9 cell line. [Method]The WS coding sequence was cloned into plasmid pFastBacTMHT B, and then transformed Escherichia coli DH10BacTM. After resistant and blue-white screening, the recombinant bacmid rBacmid/EpelWS were obtained. The rBacmid/EpelWS was transfected into Sf9 cells.[Result]The Immunofluorescence analysis showed that the target protein was expressed in the cytoplasm of the infected cells.[Conclusion]The WS was expressed successfully, it lays the foundation for further research on the biochemical characteristics of WS of the Ericerus pela.
- Ericerus pela,
- wax ester synthase,
- Construction of expression vector,
- bac-to-bac expression vector system,
- Sf9 cells lines

References

[1]	陈晓鸣, 王自力, 陈勇. 影响白蜡虫泌蜡主要气候因子及白蜡虫生态适应性分析[J]. 昆虫学报, 2007, 50(2): 136-143.
[2]	陈小平. 白蜡虫的研究进展[J]. 四川林业科技, 2007(1): 50-52.
[3]	谢映平, 薛皎亮, 张艳峰,等. 蚧虫蜡泌物的化学研究进展[J]. 昆虫知识, 2004(6): 512-518.
[4]	杨璞, 徐冬丽, 陈晓鸣, 等. 蜡酯合成途径及关键酶的研究进展[J]. 中国细胞生物学学报, 2007, 34(7): 695-703.
[5]	Pu Yang, Jia-ying Zhu, Zhong-jun et al. Transcriptome analysis of the Chinese white wax scale Ericerus pela with focus on genes involved in wax biosynthesis [J]. Plos One, 2012, 7(4): 35719.
[6]	Pu Yang, Xiao-Ming Chen, Wei-Wei Liu, et al. Transcriptome analysis of sexually dimorphic Chinese white wax scale insects reveals key differences in developmental programs and transcription factor expression[J]. Scientific Reports, 2015, (5): 8141.
[7]	Jeffrey B Cheng, David W Russell. Mammalian wax biosynthesis: II. Expression cloning of wax synthase cDNAs encoding a member of the acyltransferase enzyme family [J]. The Journal of Biological Chemistry, 2004, 279(36): 37798-37870.
[8]	Juan A. Villa, Matilde Cabezas, Fernando de la Cruz, et al. Use of limited proteolysis and mutagenesis to identify folding domains and sequence motifs critical for wax ester synthase/acylcoenzyme A: diacylglycerol acyltransferase activity [J]. Applied and Environmental Microbiology, 2014, 3(80): 1132-1141.
[9]	杨璞, 徐冬丽, 陈晓鸣,等. 蜡酯合成途径及关键酶的研究进展[J]. 中国细胞生物学学报, 2007, 34(7): 695-703.
[10]	Kalscheuer R, Luftmann H, Steinb Chel A. Synthesis of Novel Lipids in Saccharomyces cerevisiae by Heterologous Expression of an Unspecific Bacterial Acyltransferase[J]. Appl Environ Microbiol, 2004, 70: 7119-7125.
[11]	曹建斌, 范晓军, 梁爱华. 杆状病毒及其应用[J]. 科技情报开发与经济, 2007, 17(18): 131.
[12]	凌同. 昆虫杆状病毒表达系统的研究进展与应用[J]. 微生物学免疫学进展, 2014, 42(2): 70.
[13]	Rainer Kalscheuer, Alexander Steinbuchel. A novel bifunctional wax ester synthase/acyl-CoA: diacylglycerol acyltransferase mediates wax ester and triacylglycerol biosynthesis in Acinetobacter calcoaceticus ADP1 [J]. The Journal of Biological Chemistry, 2003, 278: 8075-8082.
[14]	Stefan Uthoff, Tim Stoveken, Nikolaus Weber, et al. Thio wax ester biosynthesis utilizing the unspecific bifunctional wax ester synthase/acyl coenzyme A: diacylglycerol acyltransferase of Acinetobacter sp. strain ADP1 [J]. Applied and Environmental Microbiology, 2005,(2): 790-796.
[15]	Tim Stöveken, Rainer Kalscheuer, Ursula Malkus, et al. The wax ester synthase/acyl coenzyme A:diacylglycerol acyltransferase from Acinetobacter sp. strain ADP1: characterization of a novel type of acyltransferase [J]. Journal of Bacteriology, 2005, 187(4): 76-1369.
[16]	Magdalena Miklaszewska, Adam Kawiński, Antoni Banas. Detailed characterization of the substrate specificity of mouse wax synthase [J]. Acta Biochimica Polonica, 2013(60): 209-215.
[17]	Feng-Ling Li, Xue-Min Wu, Patricia Lam, et al. Identification of the wax ester synthase/acylcoenzyme A: diacylglycerol acyltransferase WSD1 required for stem wax ester biosynthesis in Arabidopsis [J]. Plant Physiology, 2008(148): 97-107.
[18]	Shuobo Shi, Juan Octavio, Valle Rodríguez, et al. Functional expression and characterization of five wax ester synthases in Saccharomyces cerevisiae and their utility for biodiesel production [J]. Biotechnology for Biofuels, 2012 (5): 7.
[19]	Kathryn D Lardizaba, James G Metz, Tetsuo Sakamoto, et al. Purification of a jojoba embryo wax synthase, cloning of its cDNA, and production of high levels of wax in seeds of transgenic Arabidopsis [J]. Plant Physiology, 2000(122): 645-655.
[20]	Alex S. Rajangam, Satinder K. Gidda, Christian Craddock, et al. Molecular characterization of the fatty alcohol oxidation pathway for wax ester mobilization in germinated jojoba seeds [J]. Plant Physiology, 2013, 161(1): 72–80.
[21]	Pollard M R, Mckeon T, Gupta L M. Studies on biosynthesis of waxes by developing jojoba seed: II. The demonstration of wax biosynthesis by cell-free homogenates [J]. Lipids, 1979, (14): 651-662.
[22]	Brett M Barney, Rachel L Mann, Janet M. Ohlert. Identification of a residue affecting fatty alcohol selectivity in wax ester synthase [J]. Applied and Environmental Microbiology, 2013, 79(1): 396-399.
[23]	吴小锋, 曹翠平, 鲁兴萌. 昆虫杆状病毒表达系统中重组蛋白质的降解及其对策[J]. 蚕业科学, 2005, 31(4): 382-388.

Proportional views

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Get Citation

PDF

XML

Article views(2980) PDF downloads(1192) Cited by()

Proportional views

Expression of Ericerus pela Wax Ester Synthase in Spodoptera frugiperda Sf9 Cell Line

1. Research Institute of Resources Insects, Chinese Academy of Forestry, Key laboratory of Cultivating and Utilization of Resources Insects of State Forestry Administration, Kunming 650224, Yunnan, China

Received Date: 2015-11-15

Abstract: [Objective]Use Bac-to-Bac expression system to express WS in Sf9 cell line. [Method]The WS coding sequence was cloned into plasmid pFastBacTMHT B, and then transformed Escherichia coli DH10BacTM. After resistant and blue-white screening, the recombinant bacmid rBacmid/EpelWS were obtained. The rBacmid/EpelWS was transfected into Sf9 cells.[Result]The Immunofluorescence analysis showed that the target protein was expressed in the cytoplasm of the infected cells.[Conclusion]The WS was expressed successfully, it lays the foundation for further research on the biochemical characteristics of WS of the Ericerus pela.

HTML

Reference (23)

[1]	陈晓鸣, 王自力, 陈勇. 影响白蜡虫泌蜡主要气候因子及白蜡虫生态适应性分析[J]. 昆虫学报, 2007, 50(2): 136-143.
[2]	陈小平. 白蜡虫的研究进展[J]. 四川林业科技, 2007(1): 50-52.
[3]	谢映平, 薛皎亮, 张艳峰,等. 蚧虫蜡泌物的化学研究进展[J]. 昆虫知识, 2004(6): 512-518.
[4]	杨璞, 徐冬丽, 陈晓鸣, 等. 蜡酯合成途径及关键酶的研究进展[J]. 中国细胞生物学学报, 2007, 34(7): 695-703.
[5]	Pu Yang, Jia-ying Zhu, Zhong-jun et al. Transcriptome analysis of the Chinese white wax scale Ericerus pela with focus on genes involved in wax biosynthesis [J]. Plos One, 2012, 7(4): 35719.
[6]	Pu Yang, Xiao-Ming Chen, Wei-Wei Liu, et al. Transcriptome analysis of sexually dimorphic Chinese white wax scale insects reveals key differences in developmental programs and transcription factor expression[J]. Scientific Reports, 2015, (5): 8141.
[7]	Jeffrey B Cheng, David W Russell. Mammalian wax biosynthesis: II. Expression cloning of wax synthase cDNAs encoding a member of the acyltransferase enzyme family [J]. The Journal of Biological Chemistry, 2004, 279(36): 37798-37870.
[8]	Juan A. Villa, Matilde Cabezas, Fernando de la Cruz, et al. Use of limited proteolysis and mutagenesis to identify folding domains and sequence motifs critical for wax ester synthase/acylcoenzyme A: diacylglycerol acyltransferase activity [J]. Applied and Environmental Microbiology, 2014, 3(80): 1132-1141.
[9]	杨璞, 徐冬丽, 陈晓鸣,等. 蜡酯合成途径及关键酶的研究进展[J]. 中国细胞生物学学报, 2007, 34(7): 695-703.
[10]	Kalscheuer R, Luftmann H, Steinb Chel A. Synthesis of Novel Lipids in Saccharomyces cerevisiae by Heterologous Expression of an Unspecific Bacterial Acyltransferase[J]. Appl Environ Microbiol, 2004, 70: 7119-7125.
[11]	曹建斌, 范晓军, 梁爱华. 杆状病毒及其应用[J]. 科技情报开发与经济, 2007, 17(18): 131.
[12]	凌同. 昆虫杆状病毒表达系统的研究进展与应用[J]. 微生物学免疫学进展, 2014, 42(2): 70.
[13]	Rainer Kalscheuer, Alexander Steinbuchel. A novel bifunctional wax ester synthase/acyl-CoA: diacylglycerol acyltransferase mediates wax ester and triacylglycerol biosynthesis in Acinetobacter calcoaceticus ADP1 [J]. The Journal of Biological Chemistry, 2003, 278: 8075-8082.
[14]	Stefan Uthoff, Tim Stoveken, Nikolaus Weber, et al. Thio wax ester biosynthesis utilizing the unspecific bifunctional wax ester synthase/acyl coenzyme A: diacylglycerol acyltransferase of Acinetobacter sp. strain ADP1 [J]. Applied and Environmental Microbiology, 2005,(2): 790-796.
[15]	Tim Stöveken, Rainer Kalscheuer, Ursula Malkus, et al. The wax ester synthase/acyl coenzyme A:diacylglycerol acyltransferase from Acinetobacter sp. strain ADP1: characterization of a novel type of acyltransferase [J]. Journal of Bacteriology, 2005, 187(4): 76-1369.
[16]	Magdalena Miklaszewska, Adam Kawiński, Antoni Banas. Detailed characterization of the substrate specificity of mouse wax synthase [J]. Acta Biochimica Polonica, 2013(60): 209-215.
[17]	Feng-Ling Li, Xue-Min Wu, Patricia Lam, et al. Identification of the wax ester synthase/acylcoenzyme A: diacylglycerol acyltransferase WSD1 required for stem wax ester biosynthesis in Arabidopsis [J]. Plant Physiology, 2008(148): 97-107.
[18]	Shuobo Shi, Juan Octavio, Valle Rodríguez, et al. Functional expression and characterization of five wax ester synthases in Saccharomyces cerevisiae and their utility for biodiesel production [J]. Biotechnology for Biofuels, 2012 (5): 7.
[19]	Kathryn D Lardizaba, James G Metz, Tetsuo Sakamoto, et al. Purification of a jojoba embryo wax synthase, cloning of its cDNA, and production of high levels of wax in seeds of transgenic Arabidopsis [J]. Plant Physiology, 2000(122): 645-655.
[20]	Alex S. Rajangam, Satinder K. Gidda, Christian Craddock, et al. Molecular characterization of the fatty alcohol oxidation pathway for wax ester mobilization in germinated jojoba seeds [J]. Plant Physiology, 2013, 161(1): 72–80.
[21]	Pollard M R, Mckeon T, Gupta L M. Studies on biosynthesis of waxes by developing jojoba seed: II. The demonstration of wax biosynthesis by cell-free homogenates [J]. Lipids, 1979, (14): 651-662.
[22]	Brett M Barney, Rachel L Mann, Janet M. Ohlert. Identification of a residue affecting fatty alcohol selectivity in wax ester synthase [J]. Applied and Environmental Microbiology, 2013, 79(1): 396-399.
[23]	吴小锋, 曹翠平, 鲁兴萌. 昆虫杆状病毒表达系统中重组蛋白质的降解及其对策[J]. 蚕业科学, 2005, 31(4): 382-388.

Expression of Ericerus pela Wax Ester Synthase in Spodoptera frugiperda Sf9 Cell Line

Abstract

References

Proportional views

通讯作者: 陈斌, bchen63@163.com

Proportional views

Expression of Ericerus pela Wax Ester Synthase in Spodoptera frugiperda Sf9 Cell Line

HTML

Catalog

Export File

Citation

Format

Content