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Pakistan Journal of Biological Sciences

Year: 2006 | Volume: 9 | Issue: 4 | Page No.: 654-658
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Authors

Sima Rasti

Ali Haghighi

Bahram Kazemi

Mostafa Rezaian

Keywords

Research Article

Cloning and Characterization of Serine-rich Entamoeba histolytica Protein Gene from an Iranian E. histolytica Isolate

Sima Rasti, Ali Haghighi, Bahram Kazemi and Mostafa Rezaian
Entamoeba histolytica causes amebic dysentery and amebic liver abscess, placing it second only to malaria, as a cause of death resulting from parasitic protozoa. Serine-rich Entamoeba histolytica Protein gene is an immunogenic, polymorphic protein and a candidate of vaccine that was cloned and characterized in this research. An Iranian Entamoeba histolytica/Entamoeba dispar isolate was cultivated in Robinson Medium. Total genomic DNA was isolated and Entamoeba histolytica strain has been distinguished by PCR with two sets of species-specific primers and SREHP gene was amplified by Nested-PCR. The purified PCR product was cloned into pBluscript via T/A cloning method. The recombinant plasmid was screened by Rusconis test, digested with BamH1 restriction enzyme, which was approved by direct PCR and sequencing. The digested fragment of SREHP gene from recombinant plasmid was sequenced and showed a 666 base pair nucleotides as a new genotype of SREHP Entamoeba histolytica. The SREHP gene from Iranian isolate (SHR 10 IR) was different from HM1-IMSS cl.6 as well as all previously reported Entamoeba histolytica genotypes. Isolation of SREHP, expression of the gene, makes an opportunity to develop diagnostic kit, as well as determination of genetic variation of Entamoeba histolytica isolates in Iran.
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