Cloning Gene Encoding Micronema 3 (Mic3) Protein of Tachyzoite Toxoplasma Gondii Local Isolate

• Main Authors: Wayan T. Artama1, Ni Nyoman Ayu Dewi, Didik Tulus Subekti
• Format: Article
• Language: English
• Published: Universitas Gadjah Mada, Yogyakarta 2015-10-01
• Series: Indonesian Journal of Biotechnology
• Online Access: http://journal.ugm.ac.id/ijbiotech/article/view/7410

Microneme 3 (MIC3) protein tachyzoites Toxoplasma gondii is one of protein which plays an important role<br />during cell host invasion. Gene encoding MIC3 protein has been studied and it was suggested a potent vaccine<br />candidate against Toxoplasma gondii infection. The aim of this research is to clone and sequence the gene encoding<br />MIC3 protein of tachyzoites Toxoplasma gondii local isolate by amplification using polymerase chain reaction with<br />specific primers. The amplified DNA fragment was cloned into pGEM-T and transformed into E. coli XL-1 Blue by<br />heat shock method. Recombinant plasmids were isolated using alkali lysis method and analyzed by digestion<br />using restriction endonuclease enzymes PstI, HindIII, NcoI and EcoRV. The recombinant plasmids then sequenced<br />to find out the nucleotide sequence of insert gene by ABIPRISM 377 DNA Sequencer. The DNA sequence then<br />were analyzed by computer software for alignment. The result showed that transformation in E. coli XL-1 Blue by<br />pGEM-T produced one clone that was encoding MIC3 protein. Analysis of 489 bp from 5&rsquo; and 447 from 3&rsquo; of gene<br />sequence showed 97-98% homology with gene encoding for MIC3 protein of RH isolate.<br />Keywords: MIC3 protein, Toxoplasma gondii, tachyzoite, recombinant DNA