Cloning and expression a-galactosidase in order to convert B to O blood group

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Hadad Deilami A.; SALEHI M.; SHAHABI M.

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Journal Paper

Paper Information

Journal: THE SCIENTIFIC JOURNAL OF IRANIAN BLOOD TRANSFUSION Year:2023 | Volume:19 | Issue:4 Page(s): 301-312

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Information Journal Paper

Title

Cloning and expression a-galactosidase in order to convert B to O blood group

Author(s)

Hadad Deilami A. | SALEHI M. | SHAHABI M. | Issue Writer Certificate

Keywords

Blood Group

Galactosidase

E coli

Abstract

Background and Objectives The shortage of different ABO blood groups is a permanent problem that many blood transfusion centers encounter. The conversion of blood group A and B to group O is a solution to this problem. In this research we aimed to clone and express the α,-galactosidase enzyme gene to convert the blood group B to O. Materials and Methods In this experimental study, the α,-galacosidase gene from coffee bean was codon-optimized and cloned into the pBHA plasmid. After replication in E. coli, plasmid was cut with BamH1 and Nco1 restriction enzymes and the α,-galacosidase gene was purified by gel electrophoresis. The gene was then cloned into the expression vector, pET-20b. The expression construct was introduced into E. coli Rosetta 2 (DE3) and the expression of the enzyme was induced by IPTG. The presence of protein production was investigated using SDS-PAGE and Western blotting. Protein function test was also evaluated by the ability of B cells to convert to O cells. Results The presence of the recombinant protein was confirmed by SDS-PAGE and Western blot assay. The purified protein could partially convert blood group B RBCs into group O as detected by decreasing the agglutination strength with B antiserum from 4+ to less than 1+. Conclusions The production of eukaryotic proteins in a prokaryotic host is a major step in mass production. Optimization of different expression conditions is essential to achieve sufficient amount of enzyme.

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APA: Copy

Hadad Deilami, A., SALEHI, M., & SHAHABI, M.. (2023). Cloning and expression a-galactosidase in order to convert B to O blood group. THE SCIENTIFIC JOURNAL OF IRANIAN BLOOD TRANSFUSION ORGANIZATION (KHOON), 19(4 ), 301-312. SID. https://sid.ir/paper/1119964/en

Vancouver: Copy

Hadad Deilami A., SALEHI M., SHAHABI M.. Cloning and expression a-galactosidase in order to convert B to O blood group. THE SCIENTIFIC JOURNAL OF IRANIAN BLOOD TRANSFUSION ORGANIZATION (KHOON)[Internet]. 2023;19(4 ):301-312. Available from: https://sid.ir/paper/1119964/en

IEEE: Copy

A. Hadad Deilami, M. SALEHI, and M. SHAHABI, “Cloning and expression a-galactosidase in order to convert B to O blood group,” THE SCIENTIFIC JOURNAL OF IRANIAN BLOOD TRANSFUSION ORGANIZATION (KHOON), vol. 19, no. 4 , pp. 301–312, 2023, [Online]. Available: https://sid.ir/paper/1119964/en

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