Comparative method of protein expression and isolation of EBV epitope in E.coli DH5α

Nadya V.M. Anyndita; Nurul Dluha; Karimatul Himmah; Muhaimin Rifa'I

doi:10.1063/1.5012735

Comparative method of protein expression and isolation of EBV epitope in E.coli DH5α

Nadya V.M. Anyndita
, Nurul Dluha
, Karimatul Himmah
, Muhaimin Rifa'I
, Widodo^*

^*Corresponding author for this work

Faculty of Mathematics and Natural Sciences

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution › peer-review

Abstract

Epstein-Barr Virus (EBV) or human herpes virus 4 (HHV-4) is a virus that infects human B cell and leads to nasopharyngeal carcinoma (NPC). The prevention of this disease remains unsuccessful since the vaccine has not been discovered. The objective of this study is to over-produce EBV gp350/220 epitope using several methods in E.coli DH5α. EBV epitope sequences were inserted into pMAL-p5x vector, then transformed into DH5α E.coli and over-produced using 0.3, 1 and 2 mM IPTG. Plasmid transformation was validated using AflIII restriction enzyme in 0.8% agarose. Periplasmic protein was isolated using 2 comparative methods and then analyzed using SDS-PAGE. Method A produced a protein band around 50 kDa and appeared only at transformant. Method B failed to isolate the protein, indicated by no protein band appearing. In addition, any variations in IPTG concentration didn't give a different result. Thus it can be concluded that even the lowest IPTG concentration is able to induce protein expression.

Original language	English
Title of host publication	8th International Conference on Global Resource Conservation, ICGRC 2017
Subtitle of host publication	Green Campus Movement for Global Conservation
Editors	Akira Kikuchi, Chandrakhant Salunkhe, Dian Siswanto, Yoga Dwi Jatmiko, Bhupal Govinda
Publisher	American Institute of Physics Inc.
ISBN (Electronic)	9780735416000
DOIs	https://doi.org/10.1063/1.5012735
Publication status	Published - 29 Nov 2017
Event	8th International Conference on Global Resource Conservation: Green Campus Movement for Global Conservation, ICGRC 2017 - Malang, Indonesia Duration: 19 Jul 2017 → 20 Jul 2017

Publication series

Name	AIP Conference Proceedings
Volume	1908
ISSN (Print)	0094-243X
ISSN (Electronic)	1551-7616

Conference

Conference	8th International Conference on Global Resource Conservation: Green Campus Movement for Global Conservation, ICGRC 2017
Country/Territory	Indonesia
City	Malang
Period	19/07/17 → 20/07/17

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

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10.1063/1.5012735

Cite this

Anyndita, N. V. M., Dluha, N., Himmah, K., Rifa'I, M., & Widodo (2017). Comparative method of protein expression and isolation of EBV epitope in E.coli DH5α. In A. Kikuchi, C. Salunkhe, D. Siswanto, Y. D. Jatmiko, & B. Govinda (Eds.), 8th International Conference on Global Resource Conservation, ICGRC 2017: Green Campus Movement for Global Conservation Article 060002 (AIP Conference Proceedings; Vol. 1908). American Institute of Physics Inc.. https://doi.org/10.1063/1.5012735

Anyndita, Nadya V.M. ; Dluha, Nurul ; Himmah, Karimatul et al. / Comparative method of protein expression and isolation of EBV epitope in E.coli DH5α. 8th International Conference on Global Resource Conservation, ICGRC 2017: Green Campus Movement for Global Conservation. editor / Akira Kikuchi ; Chandrakhant Salunkhe ; Dian Siswanto ; Yoga Dwi Jatmiko ; Bhupal Govinda. American Institute of Physics Inc., 2017. (AIP Conference Proceedings).

@inproceedings{0327e8eee8c84b6db9f80d777ab14260,

title = "Comparative method of protein expression and isolation of EBV epitope in E.coli DH5α",

abstract = "Epstein-Barr Virus (EBV) or human herpes virus 4 (HHV-4) is a virus that infects human B cell and leads to nasopharyngeal carcinoma (NPC). The prevention of this disease remains unsuccessful since the vaccine has not been discovered. The objective of this study is to over-produce EBV gp350/220 epitope using several methods in E.coli DH5α. EBV epitope sequences were inserted into pMAL-p5x vector, then transformed into DH5α E.coli and over-produced using 0.3, 1 and 2 mM IPTG. Plasmid transformation was validated using AflIII restriction enzyme in 0.8\% agarose. Periplasmic protein was isolated using 2 comparative methods and then analyzed using SDS-PAGE. Method A produced a protein band around 50 kDa and appeared only at transformant. Method B failed to isolate the protein, indicated by no protein band appearing. In addition, any variations in IPTG concentration didn't give a different result. Thus it can be concluded that even the lowest IPTG concentration is able to induce protein expression.",

author = "Anyndita, \{Nadya V.M.\} and Nurul Dluha and Karimatul Himmah and Muhaimin Rifa'I and Widodo",

note = "Publisher Copyright: {\textcopyright} 2017 Author(s).; 8th International Conference on Global Resource Conservation: Green Campus Movement for Global Conservation, ICGRC 2017 ; Conference date: 19-07-2017 Through 20-07-2017",

year = "2017",

month = nov,

day = "29",

doi = "10.1063/1.5012735",

language = "English",

series = "AIP Conference Proceedings",

publisher = "American Institute of Physics Inc.",

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Anyndita, NVM, Dluha, N, Himmah, K, Rifa'I, M & Widodo 2017, Comparative method of protein expression and isolation of EBV epitope in E.coli DH5α. in A Kikuchi, C Salunkhe, D Siswanto, YD Jatmiko & B Govinda (eds), 8th International Conference on Global Resource Conservation, ICGRC 2017: Green Campus Movement for Global Conservation., 060002, AIP Conference Proceedings, vol. 1908, American Institute of Physics Inc., 8th International Conference on Global Resource Conservation: Green Campus Movement for Global Conservation, ICGRC 2017, Malang, Indonesia, 19/07/17. https://doi.org/10.1063/1.5012735

Comparative method of protein expression and isolation of EBV epitope in E.coli DH5α. / Anyndita, Nadya V.M.; Dluha, Nurul; Himmah, Karimatul et al.
8th International Conference on Global Resource Conservation, ICGRC 2017: Green Campus Movement for Global Conservation. ed. / Akira Kikuchi; Chandrakhant Salunkhe; Dian Siswanto; Yoga Dwi Jatmiko; Bhupal Govinda. American Institute of Physics Inc., 2017. 060002 (AIP Conference Proceedings; Vol. 1908).

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution › peer-review

TY - GEN

T1 - Comparative method of protein expression and isolation of EBV epitope in E.coli DH5α

AU - Anyndita, Nadya V.M.

AU - Dluha, Nurul

AU - Himmah, Karimatul

AU - Rifa'I, Muhaimin

AU - Widodo,

PY - 2017/11/29

Y1 - 2017/11/29

N2 - Epstein-Barr Virus (EBV) or human herpes virus 4 (HHV-4) is a virus that infects human B cell and leads to nasopharyngeal carcinoma (NPC). The prevention of this disease remains unsuccessful since the vaccine has not been discovered. The objective of this study is to over-produce EBV gp350/220 epitope using several methods in E.coli DH5α. EBV epitope sequences were inserted into pMAL-p5x vector, then transformed into DH5α E.coli and over-produced using 0.3, 1 and 2 mM IPTG. Plasmid transformation was validated using AflIII restriction enzyme in 0.8% agarose. Periplasmic protein was isolated using 2 comparative methods and then analyzed using SDS-PAGE. Method A produced a protein band around 50 kDa and appeared only at transformant. Method B failed to isolate the protein, indicated by no protein band appearing. In addition, any variations in IPTG concentration didn't give a different result. Thus it can be concluded that even the lowest IPTG concentration is able to induce protein expression.

AB - Epstein-Barr Virus (EBV) or human herpes virus 4 (HHV-4) is a virus that infects human B cell and leads to nasopharyngeal carcinoma (NPC). The prevention of this disease remains unsuccessful since the vaccine has not been discovered. The objective of this study is to over-produce EBV gp350/220 epitope using several methods in E.coli DH5α. EBV epitope sequences were inserted into pMAL-p5x vector, then transformed into DH5α E.coli and over-produced using 0.3, 1 and 2 mM IPTG. Plasmid transformation was validated using AflIII restriction enzyme in 0.8% agarose. Periplasmic protein was isolated using 2 comparative methods and then analyzed using SDS-PAGE. Method A produced a protein band around 50 kDa and appeared only at transformant. Method B failed to isolate the protein, indicated by no protein band appearing. In addition, any variations in IPTG concentration didn't give a different result. Thus it can be concluded that even the lowest IPTG concentration is able to induce protein expression.

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U2 - 10.1063/1.5012735

DO - 10.1063/1.5012735

M3 - Conference contribution

AN - SCOPUS:85038962795

T3 - AIP Conference Proceedings

BT - 8th International Conference on Global Resource Conservation, ICGRC 2017

A2 - Kikuchi, Akira

A2 - Salunkhe, Chandrakhant

A2 - Siswanto, Dian

A2 - Jatmiko, Yoga Dwi

A2 - Govinda, Bhupal

PB - American Institute of Physics Inc.

T2 - 8th International Conference on Global Resource Conservation: Green Campus Movement for Global Conservation, ICGRC 2017

Y2 - 19 July 2017 through 20 July 2017

ER -

Anyndita NVM, Dluha N, Himmah K, Rifa'I M , Widodo. Comparative method of protein expression and isolation of EBV epitope in E.coli DH5α. In Kikuchi A, Salunkhe C, Siswanto D, Jatmiko YD, Govinda B, editors, 8th International Conference on Global Resource Conservation, ICGRC 2017: Green Campus Movement for Global Conservation. American Institute of Physics Inc. 2017. 060002. (AIP Conference Proceedings). doi: 10.1063/1.5012735

Comparative method of protein expression and isolation of EBV epitope in E.coli DH5α

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