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Cloning and Sequence Analysis of Recombinant Plasmodium vivax Merozoite Surface Protein 1 (PvMSP-142 kDa) In pTZ57R/T Vector.

Mirahmadi H, Spotin A, Fallahi S, Taghipour N, Turki H, Seyyed Tabaei SJ - Iran J Parasitol (2015 Apr-Jun)

Bottom Line: Since, this region has been observed to be polymorphic among isolates of P. vivax, it is significant to survey on different regions of this antigen in various areas of the world.Thirty eight SNPs (single nucleotide polymorphism) were identified in cloned PvMSP-142 kDa gene which the mutations had localized in the 33 kDa fragment (PvMSP-133 kDa), while there was nearly no variation in the 19 kDa fragment (PvMSP-119 kDa). 2 out of 38 mutations were found as to be novel haplotypes.High similarity of cloned PvMSP-142 kDa gene in comparison to reference sequence and other sequences could be beneficial as a remarkable molecular marker for serological diagnostic kits of P. vivax in malarious neighboring countries of Iran and around the world.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences, Tehran, Iran ; Infectious Diseases and Tropical Medicine Research Center, Zahedan University of Medical Sciences, Zahedan, Iran.

ABSTRACT

Background: Carboxy-terminal 42 kDa region of Plasmodium vivax merozoite surface protein-1 is considered as an important antigen in blood stage. Since, this region has been observed to be polymorphic among isolates of P. vivax, it is significant to survey on different regions of this antigen in various areas of the world.

Methods: In the present study, the genetic diversity of cloned PvMSP-142 kDa gene from an Iranian patient is analyzed. Parasite DNA was extracted from a P. vivax - infected patient in Iran. The region of PvMSP-142 kDa was amplified by PCR, cloned into pTZ57R/T vector and then sequenced.

Results: Sequencing of cloned PvMSP-142 kDa gene clearly has a high degree of homology (95%) with reference Sal-I sequence and also with the homogeneous sequences from some studied countries (97%). Thirty eight SNPs (single nucleotide polymorphism) were identified in cloned PvMSP-142 kDa gene which the mutations had localized in the 33 kDa fragment (PvMSP-133 kDa), while there was nearly no variation in the 19 kDa fragment (PvMSP-119 kDa). 2 out of 38 mutations were found as to be novel haplotypes.

Conclusion: High similarity of cloned PvMSP-142 kDa gene in comparison to reference sequence and other sequences could be beneficial as a remarkable molecular marker for serological diagnostic kits of P. vivax in malarious neighboring countries of Iran and around the world.

No MeSH data available.


Schematic showing processing of merozoite surface protein 1 (MSP1). Panel (A) shows primary processing, and (B) shows secondary processing
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Figure 5: Schematic showing processing of merozoite surface protein 1 (MSP1). Panel (A) shows primary processing, and (B) shows secondary processing

Mentions: In research on diagnostic potential for P. vivax malaria parasite subunits, the focus is on the C-terminal region of P. vivax MSP-1. As Fig. 5 depicts, generally, MSP1 is manufactured as a big protein (∼200 kDa) which is connected to the cell membrane by a glycophosphatidylinositol (GPI) anchor in the carboxyl terminus region (11). MSP1 undergoes a series of proteolytic maturation changes at the same time as the merozoite unleashed from (RBC) and generates four polypeptide fragments of nearly 83, 30, 38, and 42 kDa from the N-terminus to C-terminus (12), which stays as interconnected parts on the cell surface by the anchored C-terminal segment (p42) (13). New RBCs are invaded rapidly by attachment of free merozoites induces second in a series cleavage of the p42 peptide to generate p33, which is shed together with the previous fragments and p19, abides by anchoring to the membrane of parasite as it invades the cell (14, 15). High plentifulness and important function on the cell surface, probably caused MSP1 to be a principal object of the host immune system and antibodies are identified on different regions of this protein (16). Antibodies that recognize the C-terminal region of Plasmodium falciparum MSP-1, inhibit the invasion of merozoites into the host erythrocytes in vitro and immunization of experimental animals with MSP-119kDa also confers protective immunity (17, 18). These findings demonstrate that MSP-142kDa is a promising candidate antigen for blood stage vaccine development and diagnostic kits.


Cloning and Sequence Analysis of Recombinant Plasmodium vivax Merozoite Surface Protein 1 (PvMSP-142 kDa) In pTZ57R/T Vector.

Mirahmadi H, Spotin A, Fallahi S, Taghipour N, Turki H, Seyyed Tabaei SJ - Iran J Parasitol (2015 Apr-Jun)

Schematic showing processing of merozoite surface protein 1 (MSP1). Panel (A) shows primary processing, and (B) shows secondary processing
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4522295&req=5

Figure 5: Schematic showing processing of merozoite surface protein 1 (MSP1). Panel (A) shows primary processing, and (B) shows secondary processing
Mentions: In research on diagnostic potential for P. vivax malaria parasite subunits, the focus is on the C-terminal region of P. vivax MSP-1. As Fig. 5 depicts, generally, MSP1 is manufactured as a big protein (∼200 kDa) which is connected to the cell membrane by a glycophosphatidylinositol (GPI) anchor in the carboxyl terminus region (11). MSP1 undergoes a series of proteolytic maturation changes at the same time as the merozoite unleashed from (RBC) and generates four polypeptide fragments of nearly 83, 30, 38, and 42 kDa from the N-terminus to C-terminus (12), which stays as interconnected parts on the cell surface by the anchored C-terminal segment (p42) (13). New RBCs are invaded rapidly by attachment of free merozoites induces second in a series cleavage of the p42 peptide to generate p33, which is shed together with the previous fragments and p19, abides by anchoring to the membrane of parasite as it invades the cell (14, 15). High plentifulness and important function on the cell surface, probably caused MSP1 to be a principal object of the host immune system and antibodies are identified on different regions of this protein (16). Antibodies that recognize the C-terminal region of Plasmodium falciparum MSP-1, inhibit the invasion of merozoites into the host erythrocytes in vitro and immunization of experimental animals with MSP-119kDa also confers protective immunity (17, 18). These findings demonstrate that MSP-142kDa is a promising candidate antigen for blood stage vaccine development and diagnostic kits.

Bottom Line: Since, this region has been observed to be polymorphic among isolates of P. vivax, it is significant to survey on different regions of this antigen in various areas of the world.Thirty eight SNPs (single nucleotide polymorphism) were identified in cloned PvMSP-142 kDa gene which the mutations had localized in the 33 kDa fragment (PvMSP-133 kDa), while there was nearly no variation in the 19 kDa fragment (PvMSP-119 kDa). 2 out of 38 mutations were found as to be novel haplotypes.High similarity of cloned PvMSP-142 kDa gene in comparison to reference sequence and other sequences could be beneficial as a remarkable molecular marker for serological diagnostic kits of P. vivax in malarious neighboring countries of Iran and around the world.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences, Tehran, Iran ; Infectious Diseases and Tropical Medicine Research Center, Zahedan University of Medical Sciences, Zahedan, Iran.

ABSTRACT

Background: Carboxy-terminal 42 kDa region of Plasmodium vivax merozoite surface protein-1 is considered as an important antigen in blood stage. Since, this region has been observed to be polymorphic among isolates of P. vivax, it is significant to survey on different regions of this antigen in various areas of the world.

Methods: In the present study, the genetic diversity of cloned PvMSP-142 kDa gene from an Iranian patient is analyzed. Parasite DNA was extracted from a P. vivax - infected patient in Iran. The region of PvMSP-142 kDa was amplified by PCR, cloned into pTZ57R/T vector and then sequenced.

Results: Sequencing of cloned PvMSP-142 kDa gene clearly has a high degree of homology (95%) with reference Sal-I sequence and also with the homogeneous sequences from some studied countries (97%). Thirty eight SNPs (single nucleotide polymorphism) were identified in cloned PvMSP-142 kDa gene which the mutations had localized in the 33 kDa fragment (PvMSP-133 kDa), while there was nearly no variation in the 19 kDa fragment (PvMSP-119 kDa). 2 out of 38 mutations were found as to be novel haplotypes.

Conclusion: High similarity of cloned PvMSP-142 kDa gene in comparison to reference sequence and other sequences could be beneficial as a remarkable molecular marker for serological diagnostic kits of P. vivax in malarious neighboring countries of Iran and around the world.

No MeSH data available.