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

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.