biostudies-literatureHanafiah MKementerian Riset Teknologi Dan Pendidikan Tinggi Republik Indonesia2085-2091https://www.ebi.ac.uk/biostudies/studies/S-EPMC7704308biostudies-literatureUnknown13(10)<h4>Aim</h4>The objective of this study was to produce recombinant protein GRA-4 (rGRA-4) of a local <i>Toxoplasma gondii</i> isolate as a candidate for a toxoplasmosis diagnosis kit in <i>Escherichia coli</i> BL21 (DE3) competent cells using pET SUMO plasmid.<h4>Materials and methods</h4>Samples used were stock <i>T. gondii</i> tachyzoites DNA from the Parasitology Laboratory, Faculty of Veterinary Medicine, Gadjah Mada University, Yogyakarta. Amplified GRA-4 polymerase chain reaction product of <i>T. gondii</i> tachyzoite DNA was cloned in the pET-SUMO TA^R cloning vector. The <i>GRA-4</i> gene from <i>T. gondii</i> local isolate was sequenced, followed by plasmid transformation, recombinant plasmid DNA isolation, and recombinant protein expression in DE3 competent cells.<h4>Results</h4>The amplification product of <i>GRA-4</i> <i>T. gondii</i> gene was 1036 bp, with 48 kDa molecular weight after expression in DE3 competent cells. An alignment of the amino acid sequence of GRA-4 from the local isolate which was cloned with GRA-4 was obtained from NCBI database and showed 99.61% homology to the predicted GRA-4 from the <i>T. gondii</i> Izatnagar isolate. Amino acid sequence of the predicted GRA-4 protein from local isolate was different at positions 19 and 304.<h4>Conclusion</h4>This research cloned rGRA-4 in pET SUMO plasmid.Veterinary worldCloning and expression of <i>Toxoplasma gondii</i> GRA-4 recombinant protein as a toxoplasmosis diagnostic kit candidate.PMC7704308019/UN11.2/LT/SP3/2019Sutriana AHanafiah MFihiruddin FHelmi TZPriyowidodo DfalseCloning and expression of <i>Toxoplasma gondii</i> GRA-4 recombinant protein as a toxoplasmosis diagnostic kit candidate.<h4>Aim</h4>The objective of this study was to produce recombinant protein GRA-4 (rGRA-4) of a local <i>Toxoplasma gondii</i> isolate as a candidate for a toxoplasmosis diagnosis kit in <i>Escherichia coli</i> BL21 (DE3) competent cells using pET SUMO plasmid.<h4>Materials and methods</h4>Samples used were stock <i>T. gondii</i> tachyzoites DNA from the Parasitology Laboratory, Faculty of Veterinary Medicine, Gadjah Mada University, Yogyakarta. Amplified GRA-4 polymerase chain reaction product of <i>T. gondii</i> tachyzoite DNA was cloned in the pET-SUMO TA^R cloning vector. The <i>GRA-4</i> gene from <i>T. gondii</i> local isolate was sequenced, followed by plasmid transformation, recombinant plasmid DNA isolation, and recombinant protein expression in DE3 competent cells.<h4>Results</h4>The amplification product of <i>GRA-4</i> <i>T. gondii</i> gene was 1036 bp, with 48 kDa molecular weight after expression in DE3 competent cells. An alignment of the amino acid sequence of GRA-4 from the local isolate which was cloned with GRA-4 was obtained from NCBI database and showed 99.61% homology to the predicted GRA-4 from the <i>T. gondii</i> Izatnagar isolate. Amino acid sequence of the predicted GRA-4 protein from local isolate was different at positions 19 and 304.<h4>Conclusion</h4>This research cloned rGRA-4 in pET SUMO plasmid.2020-01-01T00:00:00Z2020 Oct2024-12-04T10:01:45.57Z2021-02-20T10:38:19ZS-EPMC77043083328134010.14202/vetworld.2020.2085-2091