GENE CLONING OF A P35 ANTIGEN OF TOXOPLASMA GONDII

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DOROSTKAR MOGHADAM D.; KAZEMI B.; SALEHI MANSOUR; KAVAKEB P.

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Journal: Journal of Isfahan Medical School Year:2005 | Volume:23 | Issue:77 Page(s): 14-19

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Title

GENE CLONING OF A P35 ANTIGEN OF TOXOPLASMA GONDII

Author(s)

DOROSTKAR MOGHADAM D. | KAZEMI B. | SALEHI MANSOUR | KAVAKEB P. | Issue Writer Certificate

Keywords

GENE CLONINGQ4

TOXOPLASMA GONDIIQ4

P35 ANTIGENQ2

Abstract

Introduction: The use of serologic tests to show specific antibodies to the protozoan parasite TOXOPLASMA GONDII is the primary method of diagnosis. The currently available tests for the detection of IgG and IgM antibodies in infected individuals vary in their ability to detect serum antibodies. The differences observed between these serologic tests are probably due in part to the various preparations of antigen used to detect serum antibody. Due to the inherent limitations of the whole tachyzoite antigen in serologic tests, the advent of purified and tachyzoite-specific recombinant antigens obtained via molecular biology is an attractive alternative for the detection of serum antibodies.Materials and methods: A fragment of 450 bp from P35 gene of TOXOPLASMA GONDII encoding P35 tachyzoite-specific antigen was amplified by polymerase chain reaction (PCR) and then cloned into pGEM-5Z vector. The cloning was verified by Colony-PCR and Restriction Enzyme Analysis. The cloned gene was sent for sequencing and the result was evaluated using the BLAST software.Results: A 450 bp band of PCR product was seen on agarose gel and the results of Colony-PCR and Restriction Enzyme Analysis showed that the insert is cloned into the vector. DNA fragment sequence was compared to the GenBank database using BLAST software and a high score of alignment (1338) was seen between our sequence and P35 ANTIGEN sequence.Discussion: Cloning of the gene into pGEM vector has two advantages. First, we can preserve the gene for frequent use and second, with the existence of two restriction enzyme sites at each end of the gene we are able to drop the gene for sub-cloning into expression vectors. Based on sequence result there is no difference in P35 gene between RH strain existent in Iran and GenBank data and it verifies the results of previous studies from other countries witch estimate only 1% divergence at the DNA sequence level between lineages isolated from different geographical areas.

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APA: Copy

DOROSTKAR MOGHADAM, D., KAZEMI, B., SALEHI, MANSOUR, & KAVAKEB, P.. (2005). GENE CLONING OF A P35 ANTIGEN OF TOXOPLASMA GONDII. JOURNAL OF ISFAHAN MEDICAL SCHOOL (I.U.M.S), 23(77), 14-19. SID. https://sid.ir/paper/50329/en

Vancouver: Copy

DOROSTKAR MOGHADAM D., KAZEMI B., SALEHI MANSOUR, KAVAKEB P.. GENE CLONING OF A P35 ANTIGEN OF TOXOPLASMA GONDII. JOURNAL OF ISFAHAN MEDICAL SCHOOL (I.U.M.S)[Internet]. 2005;23(77):14-19. Available from: https://sid.ir/paper/50329/en

IEEE: Copy

D. DOROSTKAR MOGHADAM, B. KAZEMI, MANSOUR SALEHI, and P. KAVAKEB, “GENE CLONING OF A P35 ANTIGEN OF TOXOPLASMA GONDII,” JOURNAL OF ISFAHAN MEDICAL SCHOOL (I.U.M.S), vol. 23, no. 77, pp. 14–19, 2005, [Online]. Available: https://sid.ir/paper/50329/en

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