Publication: Molecular cloning and expression of α-Globin and β-Globin genes from crocodile (crocodylus siamensis)
Issued Date
2013-03-01
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ISSN
15734943
15723887
15723887
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2-s2.0-84876413063
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Mahidol University
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SCOPUS
Bibliographic Citation
Protein Journal. Vol.32, No.3 (2013), 172-182
Suggested Citation
Preeyanan Anwised, Thai Kabbua, Theeranan Temsiripong, Apisak Dhiravisit, Sarawut Jitrapakdee, Tomohiro Araki, Kazunari Yoneda, Sompong Thammasirirak Molecular cloning and expression of α-Globin and β-Globin genes from crocodile (crocodylus siamensis). Protein Journal. Vol.32, No.3 (2013), 172-182. doi:10.1007/s10930-013-9474-5 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/31357
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Title
Molecular cloning and expression of α-Globin and β-Globin genes from crocodile (crocodylus siamensis)
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Abstract
The first report of complete nucleotide sequences for α- and β-globin chains from the Siamese hemoglobin (Crocodylus siamensis) is given in this study. The cDNAs encoding α- and β-globins were cloned by RT-PCR using the degenerate primers and by the rapid amplification of cDNA ends method. The full-length α-globin cDNA contains an open reading frame of 423 nucleotides encoding 141 amino acid residues, whereas the β-globin cDNA contains an open reading frame of 438 nucleotides encoding 146 amino acid residues. The authenticity of both α- and β-globin cDNA clones were also confirmed by the heterologous expression in Escherichia coli (E. coli). This is the first time that the recombinant C. siamensis globins were produced in prokaryotic system. Additionally, the heme group was inserted into the recombinant proteins and purified heme-bound proteins were performed by affinity chromatography using Co2+-charged Talon resins. The heme-bound proteins appeared to have a maximum absorbance at 415 nm, indicated that the recombinant proteins bound to oxygen and formed active oxyhemoglobin (HbO2). The results indicated that recombinant C. siamensis globins were successfully expressed in prokaryotic system and possessed an activity as ligand binding protein. © 2013 Springer Science+Business Media New York.