|
Cloning and expression a-galactosidase in order to convert B to O blood group
|
A. Hadad deilami   , M. Salehi *  , M. Shahabi |
|
|
| Keywords: Key words: Blood Group, Galactosidase, E coli |
|
|
Full-Text [PDF 597 kb]
(659 Downloads)
| Abstract (HTML) (961 Views)
|
Type of Study: Research |
Subject:
Blood Transfusion
Published: 2022/12/31
|
|
|
|
|
|
|
Full-Text: (1214 Views) |
References:
- Yamamoto F. A historical overview of advances in molecular genetic/genomic studies of the ABO blood group system. Glycoconj J 2022; 39(2): 207-18.
- Gandhi MJ, Strong D, Whitaker BI, Petrisli E. A brief overview of clinical significance of blood group antibodies. Immunohematology 2018; 33(1): 4-6.
- Harmening DM, Forneris G, Tubby B. The ABO blood group system. In: Modern Blood Banking & Transfusion Practices Philadelphia, PA: FA Davis Company; 2012. p. 119-48.
- Selleng K, Jenichen G, Denker K, Selleng S, Müllejans B, Greinacher A. Emergency transfusion of patients with unknown blood type with blood group O Rhesus D positive red blood cell concentrates: a prospective, single-centre, observational study. Lancet Haematol 2017; 4(5): e218-e24.
- Shahverdi E, Moghaddam M, Talebian A, Abolghasemi H. Distribution of blood groups in the Iranian general population. Immunohematology 2016; 32(4): 135-9.
- Bakunina IY, Sova V, Nedashkovskaya O, Kuhlmann R, Likhosherstov L, Martynova M, et al. Alpha-galactosidase of the marine bacterium Pseudoalteromonas sp. KMM 701. Biochemistry (Mosc) 1998; 63(10): 1209-15.
- Golotin V, Balabanova L, Noskova YA, Slepchenko L, Bakunina IY, Vorobieva N, et al. Optimization of cold-adapted alpha-galactosidase expression in Escherichia coli. Protein Expr Purif 2016; 123: 14-8.
- Bhatia S, Singh A, Batra N, Singh J. Microbial production and biotechnological applications of α-galactosidase. Int J Biol Macromol 2020; 150: 1294-1313.
- Balabanova L, Bakunina IY, Likhatskaya G, Zvyagintseva T, Rasskazov V, editors. Glycoside hydrolases of marine bacteria are promising tools in haemotherapy. The Second International Conference on Bioenvironment, Biodiversity and Renewable Energies Bionature, Venice; 2011. p. 47-50.
- Bakunina IY, Balabanova LA, Pennacchio A, Trincone A. Hooked on α-D-galactosidases: from biomedicine to enzymatic synthesis. Crit Rev Biotechnol 2016; 36(2): 233-45.
- Hon J, Marusiak M, Martinek T, Kunka A, Zendulka J, Bednar D, et al. SoluProt: prediction of soluble protein expression in Escherichia coli. Bioinformatics 2021; 37(1): 23-8.
- Sourakov A, Paris Th. Fall Webworm, Hyphantria cunea (Drury) (Insecta: Lepidoptera: Arctiidae: Arctiinae). Available from: https://edis.ifas.ufl.edu/pdf/IN/IN878/IN878-D4sxx7ox78.pdf.
- Zhu A, Leng L, Monahan C, Zhang Z, Hurst R, Lenny L, et al. Characterization of recombinant α-galactosidase for use in seroconversion from blood group B to O of human erythrocytes. Arch Biochem Biophys 1996; 327(2): 324-9.
- Weignerová L, Filipi T, Manglová D, Křen V. Induction, purification and characterization of α-N-acetylgalactosaminidase from Aspergillus Niger. Appl Microbiol Biotechnol 2008; 79(5): 769-74.
- Olsen SK, Ota N, Kishishita S, Kukimoto-Niino M, Murayama K, Uchiyama H, et al. Crystal structure of the interleukin-15· interleukin-15 receptor α complex: insights into trans and cis presentation. J Biol Chem 2007; 282(51): 37191-204.
- Ahmed N, Afroze B, Abbas R, Khan MA, Akram M, Tahir S, et al. Method for efficient soluble expression and purification of recombinant human interleukin-15. Protein Expr Purif 2021; 177: 105746.
- Zhang YP, Gong F, Bao GQ, Gao HW, Ji SP, Tan YX, et al. B to O erythrocyte conversion by the recombinant α-galactosidasc. Chin Med J (Engl) 2007; 120(13): 1145-50.
- Gao H, Li S, Tan Y, Ji S, Wang Y, Bao G, et al. Application of α-N-acetylgalactosaminidase and α-galactosidase in AB to O red blood cells conversion. Artif Cells Nanomed Biotechnol 2013; 41(1): 32-6.
|
Sci J Iran Blood Transfus Organ 2022;19 (4): 301-312
|
Cloning and expression a-galactosidase in order to convert
B to O blood group
Hadad Deilami A.1, Salehi M.1, Shahabi M.2
1Faculty of Biological Sciences, Islamic Azad University, Tehran North Branch, Tehran, Iran
2Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran
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.
Key words: Blood Group, Galactosidase, E coli
Received: 29 May 2022
Accepted: 10 Sep 2022
Correspondence: Salehi M., PhD of Cellular & Molecular Biology. Assistant Professor of Faculty of Biological Sciences, Islamic Azad University, Tehran North Branch.
Postal Code: 1651153511 Tehran, Iran. Tel: (+9821) 77009801; Fax: (+9821) 77009848
E-mail: mitra_salehi_microbiology@yahoo.com
|
|
| Send email to the article author |
|
|
|
| Add your comments about this article |
|
|
|
|
|
.jpg)
.png)
