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

Arsyam Mawardi, Euniche R.P. F. Ramandey

Abstract


MSP1 merupakan protein yang antigenik dan paling banyak diekpresikan pada permukaan merozoit ketika menginfeksi eritrosit pasien malaria sehingga banyak dikembangkan untuk desain terapi vaksin. Proses ligasi dan transformasi gen MSP1 merupakan upaya penggandaan gen untuk menghasilkan produk yang sama ketika diekspresikan. Penelitian ini bertujuan mengkloning gen MSP1 P. falciparum  dari pasien malaria tropika di Jayapura menggunakan vektor pJET1.2/blunt dan sel kompeten E. coli DH5 sehingga didapatkan perbanyakan plasmid rekombinan yang mengandung gen MSP1. Darah yang positif mengandung P. falciparum diproses secara molekuler, diawali tahapan isolasi DNA genom, amplifikasi dengan teknik PCR, ligasi ke dalam vektor pJET1.2/blunt dan ditransformasi pada E. coli DH5α dengan metode Heat Shock Transformation, diakhiri dengan konfirmasi PCR untuk memastikan tersisipkannya gen blok 2 MSP1. Hasil penelitian menunjukkan bahwa konfirmasi keberadaan gen MSP1 dalam pJET1.2/blunt dengan PCR berhasil dilakukan. Dari total 10 koloni positif  yang ditumbuhkan dalam kultur cair, kemudian diiisolasi plasmid dan dikonfirmasi dengan PCR diperoleh pita hasil elektroferogram dengan ukuran sekitar 1049 bp yang menunjukan gen MSP1 dalam plasmid. Berdasar atas hasil tersebut, kloning gen MSP1 menggunakan vektor kloning pJET1.2/blunt dan sel kompeten E. coli DH5a telah berhasil dilakukan. 

Kata kunci: Heat Shock, ligasi, MSP1, P. falciparum, transformasi

 

Malaria-causing MSP-1 Plasmodium falciparum Ligation and Transformation in Jayapura City

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 DH5α 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 DH5α has been successfully performed. 

Key words: Heat shock, ligation, MSP-1, P. falciparum, transformation

 

 


Keywords


Heat Shock, Ligasi, MSP-1, P.falciparum, Transformasi

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DOI: https://doi.org/10.15395/mkb.v49n4.1138

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