Cloning of chikungunya virus Envelope 1 (E1) gene to pYES2/CT in Escherichia coli TOP10

F. A. Sitepu, S. Pambudi, B. S.P. Erlyandi, P. M. Tampubolon, R. Lestari

Research output: Chapter in Book/Report/Conference proceedingConference contributionpeer-review

Abstract

Chikungunya virus infection (CHIKV) causes chikungunya disease to humans which is transmitted by the Aedes mosquito. The disease causes acute fever, arthritis, and arthralgia. Chikungunya infection has been endemic in Indonesia, especially in Jambi. Detection of chikungunya disease which still uses the PCR method requires high costs. This reasoning is the basis of our research to develop a simple, fast, and accurate diagnostic test for chikungunya disease. A 7,283 bp recombinant vector carrying the CHIKV E1 (Envelope 1) gene was developed. The E1 gene measuring 1,320 bp was cloned in a pYES2/CT vector measuring 5,963 bp. The recombinant vector was then transformed into Escherichia coli TOP10 using the Transformation and Storage Solution (TSS). Transformant selection was cultured in agar medium with ampicillin antibiotics and colonies were isolated for recombinant plasmid extraction. Confirmation of the E1 gene existence was done using pYES2/CT with digestion and PCR. The results of pYES2/CT digestion electrophoresis obtained two DNA bands with a size of 1,320 and 5,963 bp, whereas the PCR method was obtained 1,640 bp and 1,320 bp. These results indicate the process of cloning the E1 CHIKV gene to pYES2/CT was successful based on confirmation with the digestion and PCR methods.

Original languageEnglish
Title of host publicationProceedings of the 5th International Symposium on Current Progress in Mathematics and Sciences, ISCPMS 2019
EditorsTerry Mart, Djoko Triyono, Tribidasari Anggraningrum Ivandini
PublisherAmerican Institute of Physics Inc.
ISBN (Electronic)9780735420014
DOIs
Publication statusPublished - 1 Jun 2020
Event5th International Symposium on Current Progress in Mathematics and Sciences, ISCPMS 2019 - Depok, Indonesia
Duration: 9 Jul 201910 Jul 2019

Publication series

NameAIP Conference Proceedings
Volume2242
ISSN (Print)0094-243X
ISSN (Electronic)1551-7616

Conference

Conference5th International Symposium on Current Progress in Mathematics and Sciences, ISCPMS 2019
CountryIndonesia
CityDepok
Period9/07/1910/07/19

Keywords

  • Chikungunya
  • E1
  • Escherichia coli TOP10
  • pYES2/CT

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