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.

1.
L. L.
Coffey
,
A.
Failloux
and
S. C.
Weaver
,
Viruses
6
,
4628
4663
(
2014
).
2.
O.
Schwartz
and
M. L.
Albert
,
Nat. Rev. Microbiol.
8
,
491
500
(
2010
).
3.
H.
Harapan
 et al 
BMC Infect. Dis.
19
, (
2019
).
4.
L. A.
Silva
and
T. S.
Dermody
,
J. Clin. Invest
127
,
737
749
(
2017
).
5.
F. J.
Burt
,
M. S.
Rolph
,
N. E.
Rulli
,
S.
Mahalingam
and
M. T.
Heise
,
The Lancet
379
,
662
671
(
2012
).
6.
B.
Pastorino
 et al,
J. Virol. Methods
124
,
65
71
(
2005
).
7.
M.
Kumar
,
A. B.
Sudeep
and
V. A.
Arankalle
,
Vaccine
30
,
6142
6149
(
2012
).
8.
H.
Kosasih
 et al,
PLoS Neglect. Trop. D.
7
, (
2013
).
9.
J.
Shukla
 et al,
Diagn. Micr. Infec. Dis.
65
,
142
149
(
2009
).
10.
K. K.
Yathi
,
J. M.
Joseph
,
S.
Bhasker
,
R.
Kumar
and
M.
Chinnamma
,
J. Immunol. Methods
372
,
171
176
(
2011
).
11.
R. T.
Sasmono
 et al,
Am. J. Trop. Med. Hyg.
97
,
1393
1398
(
2017
).
12.
B.
Cho
 et al,
Yonsei Med. J.
49
,
828
835
(
2008
).
13.
M. S. H.
Sum
,
A.
Andrew
and
M. A.
Maling
,
J. Biochem. Microbiol. Biotechn.
3
,
26
29
(
2015
).
14.
C. T.
Chung
,
S. L.
Niemela
and
R. H.
Miller
,
Proc. Natl. Acad. Sci.
86
,
2172
2175
(
1989
).
15.
V.
Narita
,
R. M.
Widyanto
,
S.
Pambudi
and
T. M.
Sudiro
,
Makara J. Sci.
15
,
173
178
(
2011
).
16.
J. F.
Sambrook
and
D. W.
Russell
,
Molecular Cloning: A Laboratory Manual
, 3
rd
Ed.
(
Cold Spring Harbor Laboratory Press
,
New York
,
2001
).
17.
R. D.
Gietz
and
R. H.
Schiestl
,
Nature Protocols
2
,
31
34
(
2007
).
18.
Invitrogen, pYES2/CT, pYES3/CT, and pYC2/CT Yeast expression vectors with C-terminal tags and auxotrophic selection markers
, (
2009
), available at http://2012.igem.org/wiki/images/d/da/PYES2%2C_pYES3_manual.pdf
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