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Published on: July 12, 2013
[Cloning DNA Vaccine Candidate SAG1 Gene of Toxoplasma gondii]
Hüsniye Lalek1, Esra Gürbüz, Serkan Karaca
1Erciyes Üniversitesi Tıp Fakültesi, Parazitoloji Anabilim Dalı, Kayseri, Türkiye. salihkuk@hotmail.com.
Objective:
Toxoplasma gondii, which is observed in our country and worldwide, can cause mortality and is an important public health problem because of engaging babies from pregnant women. An effective vaccine against toxoplasmosis has not yet been developed. SAG1 protein is released from the bradyzoites and tachyzoites stages of T. gondii and is important at the pathogenesis of the disease. This study aimed to clone a promising DNA vaccine candidate, SAG1 gene, of T. gondii.
Methods:
T. gondii genomic DNA was isolated from tachyzoites of T. gondii. SAG1 gene was amplified with specific primers and then cloned into the pJET1.2 vector. Recombinant plasmids were transformed into Escherichia coli cells. The presence of recombinant plasmids was determined by polymerase chain reaction (PCR) screening. Following the purification of the recombinant plasmid from positive colonies, cloning was confirmed by PCR, restriction enzyme assays, and DNA sequence analysis.
Results:
After PCR with SAG1 gene-specific primers, 1010-bp PCR products were obtained. Recombinant plasmid, which was transformed into competent E. coli cells, was verified by PCR screening. Moreover, PCR verified that the recombinant plasmids contained the SAG1 gene. The DNA sequence was analyzed, and the DNA sequence was obtained.
Conclusion:
One of the promising DNA vaccine candidates against toxoplasmosis, SAG1 gene, has been cloned.
