Berkala Penelitian Hayati
Vol 11 No 1 (2005): December 2005

VERIFIKASI cDNA T29 SEBAGAI KANDIDAT GEN PENGKODE PROTEIN Toxoplasma gondii DENGAN METODE SDS PAGE

Ira Djajanegara (P3T Bioindustri BPPT)
Wayan Artama (PAU-Bioteknologi UGM)
Retno Lestari (Departemen Biologi UI)
Sabar Pambudi (P3T Farmasi Medika BPPT)



Article Info

Publish Date
05 Feb 2012

Abstract

The process of cDNA construction from mRNA isolated from Toxoplasma gondii has been done. There were 7 candidates cDNA which one of them is called T29. Since Toxoplasma gondii is the cause of toxoplasmosis infection, cloning the gene encoding protein from this parasite provides an important tool for developing diagnostic kit for detection of toxoplasmosis. Digestion of the cDNA T29 with EcoRI which is the restriction site where the cDNA was inserted yielded a 1.862 bp fragment. The fragment was subcloned into E. coli expression vector pMal-p2x and transformed into E.coli strain TB1. Colonies of TB1 were grown on ampicillin plates and the recombinant plasmid was extracted using the standard procedure. The plasmid was digested using EcoRI and PstI, checked by PCR amplification using malE and M13/pUC primers. The recombinant plasmid was expressed in TB1 and the protein extracted was ran in SDS PAGE to observe the presence of the expressed protein. Based on the data from this experiment, there was no expression result of the expressed cDNA which was confirm by the PCR result. Therefore, it was concluded that cDNA T29 was not carrying the gene coding for protein from parasite Toxoplasma gondii.

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Journal Info

Abbrev

jurnal

Publisher

Subject

Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Immunology & microbiology Materials Science & Nanotechnology Medicine & Pharmacology

Description

Berkala Penelitian Hayati is a half yearly international peer reviewed, an open access life science journal. The journal was published by The East Java Biological Society and formerly used the Indonesian language. The first edition of this journal is Vol 1 No 1 in June 1995. It was accredited by ...