Kloning Dan Ekspresi Cdna Yang Menjadi ProTein Membran Takizoit Toxoplasma gondii Isolat Bogor = Cloning and Expression of cDNA Encoding Membrane Protein Tachyzoite of Toxoplasma gondii Bogor Isolate

ABSTRACT Diagnosis of toxoplasmosis, a diseases causes by obligatory intracellular protozoan parasite Toxoplasma gondii generally relies on serological methods. However, immunodiagnosis might be indistinct in immunocompromised patients, during pregnancy or in patient with congenital toxoplasmosis. Therefore, cloning and expression of cDNA encoding protein tachyzoite of Toxoplasma gondii for molecular diagnosis purposes was the aim of this research. Total ribonucleic acid (RNA) and messenger RNA was isolated from tachyzoites that was grown up in Swiss strain mice. Messenger RNA was isolated from total RNA using polyATtract®mRNA Isolation Systems (Promega) and synthesis of cDNA using Riboclone®cDNA Synthesis Systems (Promega). Complementary DNA was ligated with EcoR I adaptor using Ribocicmea EcoR I Adaptor Ligation System (Promega). Bacteriophage of 2 gt11 ligated with cDNA was transvected in E. coli Y1090. Then recombinant plaque was analyzed using polyclonal antibodies against membrane protein tachyzoite of Toxoplasma gondii. Over expression recombinant was analyzed by dot blot technique to identify recombinant with cDNA encoding membrane protein tachyzoite of Toxoplasma gondi. Transvection in E. coli Y1090 by 4t11 produce seven recombinant plaques encoding membrane protein tachyzoites of Toxoplasma gondii. Key words: Toxoplasma gondii, tachyzoite, cDNA, membrane protein