Ligasi dan Transformasi Gen MSP1 Plasmodium falciparum Penyebab Malaria di Kota Jayapura

Abstract

MSP1 is the most antigenic and expressed protein on merozoite surface when it infects the erythrocytes of malaria patients which leads to its use for vaccine therapy design development. The ligation and transformation process of the MSP1 gene is a gene duplication attempt for producing the same product during expression. This study aimed to clone P. falciparum MSP-1 gene from tropical malaria patients in Jayapura using pJET1.2/blunt vectors and E. coli DH5a competent cells, to get the recombinant plasmid propagation of MSP1 gene. Blood that was positive for P. falciparum was molecularly processed, starting with genomic DNA isolation and then followed by PCR amplification, ligation into pJET1.2/blunt vector, and transformation into E. coli DH5a using the heat shock transformation method. The process was ended with PCR confirmation to confirm MSP1 gene insertion. The results showed that the presence of the MSP1 gene in pJET1.2/blunt was successfully confirmed through PCR. From a total of 10 positive colonies grown in liquid culture, plasmid was isolated. Electropherogram result presented bands of about 1049bp, indicating the presence of the MSP1 gene in plasmid. Hence, MSP1 gene cloning using pJET1.2/ blunt cloning vector and competent cell E. coli DH5a has been successfully performed.